Provided herein are compositions, methods and uses involving antibodies that specifically bind to ALK, a receptor tyrosine kinase, and modulate the expression and/or activity of ALK. Also provided are uses and methods for treating, e.g., managing, disorders, such as cancer.

Description

79 parts

›This application claims the benefit of U.S. Provisional…

This application claims the benefit of U.S. Provisional Application No. 62/210,705, filed Aug. 27, 2015, the disclosure of which is incorporated by reference herein in its entirety.

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This application incorporates by reference a Sequence Listing submitted with this application as text file entitled “Sequence_Listing_12638_115_228.txt” created on Aug. 26, 2016 and having a size of 225,594 bytes.

1. FIELD

Provided herein are compositions, methods and uses involving antibodies that specifically bind to ALK, a receptor tyrosine kinase, and modulate the expression and/or activity of ALK for treating, e.g., managing, disorders, such as cancer.

2. BACKGROUND

Anaplastic Lymphoma Kinase (ALK) is a receptor tyrosine kinase (RTK) involved in neurogenesis during embryonic development. ALK is transiently expressed in specific regions of the central and peripheral nervous system, for example the mid-brain, thalamus, olfactory bulb, and peripheral ganglia. This expression is essential and is highest in the neonatal brain. Expression is maintained at low levels in the adult brain.

ALK has also been shown to be expressed in cells exhibiting a number of disease states, such as cells involved in cancer and other hyperproliferative and neoplastic disorders. Overexpression and/or constitutive activation (such as by fusion proteins) of ALK has also been associated with oncogenic growth and the formation of tumors. In addition, aberrant ALK signaling has been implicated in driving several types of cancer. Genomic translocations resulting in the fusion of the ALK kinase domain with the oligomerization region of intracellular proteins (e.g., EML4-ALK, NPM-ALK, etc.) have been identified in approximately 5% of non-small cell lung cancers, and approximately 60% of anaplastic large-cell lymphomas. Furthermore, ALK activation through overexpression, somatic mutation, or germ-line mutation occurs in approximately 15% of neuroblastoma cases, the most common extra-cranial tumor type in children.

3. SUMMARY

In one aspect, provided herein are antigen-binding agents, e.g., antibodies, which specifically bind to an extracellular domain (ECD) of ALK such as human ALK. In one aspect, provided herein are antibodies (e.g., monoclonal antibodies), including antigen-binding fragments thereof, which specifically bind to the extracellular domain (ECD) of ALK, such as human ALK. In another specific aspect, such anti-ALK antibodies specifically bind to an ECD of human ALK (e.g., the NTR, MAM1, LDLa, MAM2 or EGF domains of ALK-ECD) and inhibit phosphorylation of ALK. In a specific aspect, such anti-ALK antibodies specifically bind to an ECD of human ALK (e.g., the NTR, MAM1, LDLa, MAM2 or EGF domains of human ALK-ECD) present on the surface of a cell, and is internalized by the cell. Also provided herein are polynucleotides and vectors comprising sequences encoding such antibodies, cells (e.g., host cells) comprising such polynucleotides or vectors, and compositions, reagents and kits comprising such antibodies. In another aspect, provided herein are methods for modulating ALK activity or ALK expression levels, diagnostic methods and uses, and therapeutic methods and uses of such anti-ALK antibodies.

In one aspect, provided herein is an antibody, e.g., a monoclonal antibody, or antigen-binding fragment thereof that binds to the ECD of human ALK, wherein the antibody or antigen-binding fragment thereof:

(a) inhibits ALK activity in cells as determined by inhibition of phosphorylation of ALK;

(b) binds to the ECD of human ALK present on a cell, and is internalized by the cell;

(d) inhibits tumor cell proliferation or tumor growth.

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(a) a light chain variable region (VL) comprising VL complementary determining region 1 (CDR1), VL CDR2, and VL CDR3 of any one of antibodies Ab320-Ab332 and Ab351-Ab446, for example, as set forth in Table 1; and

(b) a heavy chain variable region (VH) comprising VH complementary determining region 1 (CDR1), VH CDR2, and VH CDR3 of any one of antibodies Ab320-Ab332 and Ab351-Ab446, for example as set forth in Table 2.

Also provided herein is a method of making an antibody or an antigen-binding fragment thereof which specifically binds to the ECD of human ALK, comprising culturing a cell provided herein to express the antibody or antigen-binding fragment.

Also provided herein is a method of making an antibody or an antigen-binding fragment thereof which specifically binds to the ECD of human ALK, comprising expressing a polynucleotide provided herein.

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a light chain variable region (VL) comprising:

(a) a VL complementarity determining region (CDR) 1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13); (b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and (c) a VL CDR3 comprising the amino acid sequence of QX 1 YNSYPYMX 2 T (SEQ ID NO:468), wherein X 1 is Q or R and X 2 is Y or F; and

(ii) a heavy chain variable region (VH) comprising:

(a) a VH CDR1 comprising the amino acid sequence X 3 YWMH (SEQ ID NO:469), wherein X 3 is N or S; (b) a VH CDR2 comprising the amino acid sequence of YIX 4 PSSGYTKYNQKFKD (SEQ ID NO:470), wherein X 4 is N or K; and (c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18).

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising:

(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising:

(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of SYWMH (SEQ ID NO:10);

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(b) a VH CDR2 comprising the amino acid sequence of YIKPSSGYTKYNQKFKD (SEQ ID NO:34); and

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of SYWMH (SEQ ID NO:10);

(b) a VH CDR2 comprising the amino acid sequence of YINPSSGYTKYNQKFKD (SEQ ID NO:17); and

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of NYWMH (SEQ ID NO:16);

(b) a VH CDR2 comprising the amino acid sequence of YINPSSGYTKYNQKFKD (SEQ ID NO:17); and

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13); (b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and (c) a VL CDR3 comprising the amino acid sequence of QQYNSYPYMYT (SEQ ID NO:15); and

(ii) a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of SYWMH (SEQ ID NO:10); (b) a VH CDR2 comprising the amino acid sequence of YIKPSSGYTKYNQKFKD (SEQ ID NO:34); and (c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18).

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(ii) a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of SYWMH (SEQ ID NO:10); (b) a VH CDR2 comprising the amino acid sequence of YINPSSGYTKYNQKFKD (SEQ ID NO:17); and (c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18).

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(ii) a VH comprising:

(a) a VH CDR1 comprising the amino acid sequence of NYWMH (SEQ ID NO:16); (b) a VH CDR2 comprising the amino acid sequence of YINPSSGYTKYNQKFKD (SEQ ID NO:17); and (c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18).

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(ii) a VH comprising:

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(ii) a VH comprising:

In certain aspects, the antibody or antigen-binding fragment thereof comprises:

(i) a VL comprising:

(ii) a VH comprising:

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 399, and a VH comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 400.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 407, and a VH comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 408.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 411, and a VH comprising an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO: 412.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 80% identical to any one of the amino acid sequences of SEQ ID NOs: 421-428, and a VH comprising an amino acid sequence at least 80% identical to any one of the amino acid sequences of SEQ ID NOs: 429-440.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 399, and a VH comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 400.

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In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 407, and a VH comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 408.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 411, and a VH comprising an amino acid sequence at least 85% identical to the amino acid sequence of SEQ ID NO: 412.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 85% identical to any one of the amino acid sequences of SEQ ID NOs: 421-428, and a VH comprising an amino acid sequence at least 85% identical to any one of the amino acid sequences of SEQ ID NOs: 429-440.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 399, and a VH comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 400.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 407, and a VH comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 408.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 411, and a VH comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 412.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 90% identical to any one of the amino acid sequences of SEQ ID NOs: 421-428, and a VH comprising an amino acid sequence at least 90% identical to any one of the amino acid sequences of SEQ ID NOs: 429-440.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 399, and a VH comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 400.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 407, and a VH comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 408.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 411, and a VH comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 412.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising an amino acid sequence at least 95% identical to any one of the amino acid sequences of SEQ ID NOs: 421-428, and a VH comprising an amino acid sequence at least 95% identical to any one of the amino acid sequences of SEQ ID NOs: 429-440.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising SEQ ID NO: 399 and a VH comprising SEQ ID NO: 400.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising SEQ ID NO: 407 and a VH comprising SEQ ID NO: 408.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising SEQ ID NO: 411 and a VH comprising SEQ ID NO: 412.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a VL comprising any one of the amino acid sequences of SEQ ID NOs: 421-428 and a VH comprising any one of the amino acid sequences of SEQ ID NOs: 429-440.

In certain aspects, the antibody or antigen-binding fragment thereof comprises the VL CDR1-3 and the VH CDR1-3 of any one of Ab320-Ab332, Ab353, Ab354, Ab358, Ab362, Ab423, or Ab435.

In certain aspects, the antibody or antigen-binding fragment thereof comprises the VL CDR1-3 and the VH CDR1-3 of any one of Ab320-Ab332, or Ab351-Ab446.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a heavy chain constant region. In certain aspects, the heavy chain constant region is a human IgG constant region. In specific aspects, the heavy chain constant region is a human IgG 1 constant region.

In certain aspects, the antibody or antigen-binding fragment thereof comprises a light chain constant region selected from the group consisting of a human kappa constant region and a human lambda constant region.

In certain aspects, the antibody or antigen-binding fragment thereof is a human antibody, a humanized antibody, a chimeric antibody, a recombinant antibody, a multispecific antibody, or an antigen-binding fragment thereof. In certain aspects, the antigen-binding fragment is an Fv, Fab, F(ab′)2, Fab′, dsFv, scFv, or sc(Fv)2. In specific aspects, the antibody or antigen-binding fragment thereof is a monoclonal antibody or antigen-binding fragment thereof. In certain aspects, the antibody or antigen-binding fragment thereof is an antibody.

In certain aspects, provided herein are compositions comprising an antibody or antigen-binding fragment thereof provided herein and a pharmaceutically acceptable carrier.

In certain aspects, provided herein are nucleic acids comprising a sequence encoding an antibody or antigen-binding fragment provided herein. In certain aspects, provided herein is a vector comprising a nucleic acid encoding an antibody or antigen-binding fragment provided herein. In certain aspects, provided herein is a cell comprising a nucleic acid encoding an antibody or antigen-binding fragment provided herein.

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In certain aspects, provided herein is a method of making an antibody or antigen-binding fragment thereof provided herein, comprising (a) culturing a cell comprising a nucleic acid encoding the antibody or antigen-binding fragment; and (b) isolating the antibody or antigen-binding fragment thereof.

In certain aspects, provided herein is a method of inhibiting the proliferation of a cell expressing ALK, said method comprising contacting the cell with an antibody or antigen-binding fragment provided herein.

In certain aspects, provided herein is a method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof provided herein. In certain aspects, the cancer is selected from the group consisting of anaplastic large cell lymphoma, inflammatory myofibroblastic tumors, lung cancer, for example, small cell lung carcinoma or non-small cell lung cancer, e.g., adenocarcinoma, such as bronchioalveolar carcinoma, squamous cell carcinoma or large-cell carcinoma, diffuse large B-cell lymphoma, squamous cell carcinoma, breast carcinoma, melanoma, pancreatic cancer, B-cell non-Hodgkin's lymphoma, thyroid carcinoma, retinoblastoma, Ewing sarcoma, prostate cancer, colon cancer, colorectal cancer, glioblastoma, rhabdomyosarcoma, ovarian cancer, head and neck cancer, e.g., head and neck squamous cell carcinomas, medulloblastoma and neuroblastoma. In specific aspects, the cancer is glioblastoma or neuroblastoma. In certain aspects the cancer comprises cells expressing ALK.

In specific aspects, the cancer comprises cells comprising an ALK mutation. In particular embodiments, the ALK mutation comprises a mutation of ALK amino acid residue 1174 (SEQ ID NO:467). In specific embodiments, the ALK mutation comprises a cysteine, isoleucine, leucine, serine, or valine amino acid substitution at ALK amino acid residue Phe1174 of SEQ ID NO:467. In specific embodiments, the ALK mutation comprises a leucine substitution at ALK amino acid residue Phe1174 of SEQ ID NO:467. In particular embodiments, the ALK mutation comprises a glutamine amino acid substitution at ALK amino acid residue Arg1275 of SEQ ID NO:467. In particular embodiments, the ALK mutation comprises a mutation of one or more of the following residues of SEQ ID NO:467: Pro36, Pro157, Va1198, Arg 259, Gly640, Leu684, Gly718, Met770, Asp993, Arg1060 (e.g., Arg1060His), Asp1091, Gly1128, Thr1151 (e.g., Thr1151Met), Met1166 (e.g., Met1166Arg), Ile1170 (e.g., Ile1170Asn or Ser), Ile1171 (e.g., Ile1171Asn), Phe1174, Arg1192, Leu1196 (e.g., Leu1196Met), Ala1200 (e.g., Ala1200Val), Leu1204 (e.g., Leu1204Phe), Glu1407, Glu1433, Leu1240 (e.g., Leu1240Val), Phe1245 (e.g., Phe1245Cys, Ile, Leu, or Val), Ile1250, Arg1464, Asp1270 (e.g., Asp1270Gly), Arg1275 (e.g., Arg1275Gln or Leu), Tyr1278 (e.g., Tyr1278Ser), Gly1286 (e.g., Gly1286Arg), Thr1343 (e.g, Thr1343Ile), Gly1494, or Ala1553. In particular embodiments, the ALK mutation is an ALK fusion gene, for example, an echinoderm microtubule-associated protein-like 4 (EML4)-ALK fusion gene, a nucleophosmin (NPM)-ALK fusion gene, or a tropomyosin 3 (TPM3)-ALK fusion gene. In specific embodiments, a EML4-ALK fusion gene is associated with lung cancer, for example, non-small cell lung cancer. In specific embodiments, a NPM-ALK fusion gene and/or a TPM3-ALK fusion gene is associated with anaplastic large-cell lymphoma.

In certain aspects, the subject being treated is a pediatric subject. For example, in certain aspects, provided herein is a method for treating cancer in a pediatric subject, comprising administering to the pediatric subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof provided herein. In certain aspects, the cancer is glioblastoma, rhabdomyosarcoma, neuroblastoma, or medulloblastoma. In specific aspects, the cancer comprises cells comprising an ALK mutation. In particular embodiments, the ALK mutation comprises a mutation of ALK amino acid residue 1174 (SEQ ID NO:467). In specific embodiments, the ALK mutation comprises a cysteine, isoleucine, leucine, serine, or valine amino acid substitution at ALK amino acid residue Phe1174 of SEQ ID NO:467. In specific embodiments, the ALK mutation comprises a leucine substitution at ALK amino acid residue Phe1174 of SEQ ID NO:467. In particular embodiments, the ALK mutation comprises a glutamine amino acid substitution at ALK amino acid residue Arg1275 of SEQ ID NO:467. In particular embodiments, the ALK mutation comprises a mutation of one or more of the following residues of SEQ ID NO:467: Pro36, Pro157, Va1198, Arg 259, Gly640, Leu684, Gly718, Met770, Asp993, Arg1060 (e.g., Arg1060His), Asp1091, Gly1128, Thr1151 (e.g., Thr1151Met), Met1166 (e.g., Met1166Arg), Ile1170 (e.g., Ile1170Asn or Ser), Ile1171 (e.g., Ile1171Asn), Phe1174, Arg1192, Leu1196 (e.g., Leu1196Met), Ala1200 (e.g., Ala1200Val), Leu1204 (e.g., Leu1204Phe), Glu1407, Glu1433, Leu1240 (e.g., Leu1240Val), Phe1245 (e.g., Phe1245Cys, Ile, Leu, or Val), Ile1250, Arg1464, Asp1270 (e.g., Asp1270Gly), Arg1275 (e.g., Arg1275Gln or Leu), Tyr1278 (e.g., Tyr1278Ser), Gly1286 (e.g., Gly1286Arg), Thr1343 (e.g, Thr1343Ile), Gly1494, or Ala1553. In particular embodiments, the ALK mutation is an ALK fusion gene, for example, an echinoderm microtubule-associated protein-like 4 (EML4)-ALK fusion gene, a nucleophosmin (NPM)-ALK fusion gene, or a tropomyosin 3 (TPM3)-ALK fusion gene. In specific embodiments, a EML4-ALK fusion gene is associated with lung cancer, for example, non-small cell lung cancer. In specific embodiments, a NPM-ALK fusion gene and/or a TPM3-ALK fusion gene is associated with anaplastic large-cell lymphoma.

In certain aspects, provided herein is a method of treating neurofibromatosis in a subject, comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof provided herein. In certain aspects, the subject is a pediatric subject.

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In certain aspects, provided herein is a kit comprising an antibody or antigen-binding fragment thereof provided herein.

In certain aspects, provided herein is a method of diagnosing a ALK-expressing cancer in a patient, wherein the method comprises the steps of:

(a) contacting a biological sample from the patient with an antibody or antigen-binding fragment provided herein;

(b) detecting binding of the antibody or antigen-binding fragment to ALK to determine an ALK protein level in the biological sample from the patient; and

In certain aspects, provided herein is a method of monitoring the ALK protein level during treatment of a ALK-expressing cancer in a patient, wherein the method comprises the steps of:

(a) administering to the patient an antibody or antigen-binding fragment provided herein;

(b) contacting a biological sample from the patient with the antibody or antigen-binding fragment;

(c) detecting binding of the antibody or antigen-binding fragment to ALK to determine an ALK protein level in the biological sample from the patient; and (d) comparing the ALK protein level with a standard ALK protein level.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts an exemplary amino acid sequence of human ALK (SEQ ID NO: 467), wherein amino acid residues 1-20 comprise the signal sequence.

FIG. 2 depicts the ALK domain arrangement and antigens used for animal immunization.

FIG. 3 depicts the apparent affinities (K D ) of ALK antibodies for ALK expression on the surface of NB1 cells. K D was determined by flow cytometry. Titration of each antibody on NB1 cells for 2-4 hours at 4° C. was followed by incubation with a fluorescently-labeled secondary antibody. The normalized mean fluorescent intensity (WI) shown as “% Max binding” was plotted as a function of antibody concentration.

FIG. 4A depicts a Western blot depicting NB1 cells grown in media with reduced serum and treated with 30-100 nM of each antibody or the ALK inhibitor crizotinib for 1-2 hours. Lysed cells were subjected to SDS-PAGE, transferred to nitrocellulose and immunoblotted with antibodies against total, and phospho-specific ALK, AKT, and MAPK. FIG. 4B depicts an ELISA of NB1 cells grown in media with reduced serum and treated with 30-100 nM of each antibody for 1-2 hours. Cell lysates were incubated on ELISA plates with an anti-ALK antibody, followed by incubation with a pan phosphotyrosine antibody coupled to HRP.

FIG. 5 depicts a Western blot of a panel of ALK antibodies. 24 hour treatment of NB1 cells with 100 nM of anti-ALK mAbs resulted in differences in ALK turnover (as measured by total protein levels).

FIG. 6 depicts an ELISA showing the effect of ALK antibodies on the proliferation of NB1 cells. NB1 cells grown in media with reduced serum were treated with 30-100 nM of each antibody for 1-2 hours. Cell lysates were incubated on ELISA plates with an anti-ALK antibody, followed by incubation with a pan phosphotyrosine antibody coupled to HRP.

FIG. 7A depicts the effect of the titration of crizotinib, a small molecule ALK tyrosine kinase inhibitor, on the killing of NB1 cells. FIG. 7B depicts the co-titration of crizotinib with ALK inhibitory antibodies (Ab326 and Ab327) and ALK agonistic antibodies (Ab323 and Ab324).

FIGS. 8A-B depict the in vivo antitumor activity of the Ab326 anti-ALK antibody in a patient-derived (PDX) neuroblastoma ALK F1174L tumor model compared with control IgG1 (A) and a western blot of tumor homogenates harvested from the patient-derived tumor model showing relative levels of total ALK and phosphorylated ALK in Ab326-treated and control IgG1-treated mice (B). GAPDH is included in the western blot of FIG. 8B as a control for protein level.

FIG. 9 depicts a sequence alignment of ALK species orthologs: human=hALK (SEQ ID NO: 519; see also SEQ ID NO: 467); mouse=mALK (SEQ ID NO: 517); rat=rALK (SEQ ID NO: 518). Boxed residues differ between rat and mouse ALK. Bold residues indicate the C-terminal ALK ECD antigen, used as an immunogen.

5. DETAILED DESCRIPTION

In one aspect, provided herein are antibodies (“ALK antibody” or “anti-ALK antibody), e.g., monoclonal antibodies, and antigen-binding fragments thereof, that specifically bind to an ALK polypeptide (e.g., an ECD of human ALK). In certain embodiments, such antibodies or antigen-binding fragments modulate ALK activity (e.g., ALK signaling) and/or ALK expression. In a specific embodiment, such antibodies or antigen-binding fragments, which specifically bind to an ECD of human ALK (e.g., the NTR, MAM1, LDLa, MAM2 or EGF domains of human ALK-ECD), bind to cell surface ALK and are internalized by the cell, and/or inhibit ALK activity (e.g., inhibit phosphorylation of ALK, induce ALK degradation, and/or inhibit tumor cell proliferation or tumor growth). Also provided are isolated nucleic acids (polynucleotides), such as complementary DNA (cDNA), encoding such antibodies, and antigen-binding fragments thereof. Further provided are vectors (e.g., expression vectors) and cells (e.g., host cells) comprising nucleic acids (polynucleotides) encoding such antibodies or antigen-binding fragments thereof. Also provided are methods of making such antibodies, antigen-binding fragments, and cells. In other aspects, provided herein are methods and uses for modulating ALK expression or ALK activity (e.g., inhibiting ALK expression or ALK activity), or treating or managing certain conditions or disorders described herein, such as treating or managing cancer. Related compositions (e.g., pharmaceutical compositions), kits, and diagnostic methods are also provided.

As used herein, the terms “ALK” or “ALK receptor” or “ALK polypeptide” refer to mammalian ALK, e.g., human ALK, including, for example, native ALK, ALK isoforms, and ALK fusion polypeptides resulting from genomic rearrangements, e.g., translocations. Native ALK is a transmembrane polypeptide that comprises an extracellular domain (ECD), a transmembrane domain, and a cytoplasmic domain. The ALK ECD comprises a basic N-terminal domain, two Meprin/A5/protein tyrosine phosphatase Mu (MAM) domains that flank a low-density lipoprotein class A (LDLa) domain, and a membrane-proximal glycine-rich domain. In a specific embodiment, the ALK is a human ALK of the amino acid sequence of SEQ ID NO: 467. GenBank™ accession numbers NP_004295.2 provides an exemplary human ALK amino acid sequence. GenBank™ accession number NM_004304.4 provides an exemplary human ALK nucleic acid. Unless otherwise specified herein, references to particular amino acid residues of ALK correspond to the amino acid residues of the human form of ALK set forth in SEQ ID NO: 467 and reproduced below:

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The ALK extracellular domain corresponds to amino acids 21-1038, the N-terminal domain corresponds to amino acids 21-263, the first MAM domain (MAM1) corresponds to amino acids 264-427, the LDLa domain corresponds to amino acids 437-473), the second MAM domain (MAM2) corresponds to amino acids 478-636, the glycine-rich domain corresponds to amino acids 816-940, the transmembrane domain corresponds to amino acids 1039-1059, the intracellular domain corresponds to amino acids 1060-1620, with the kinase domain corresponding to amino acids 1116-1392; the immature ALK sequence also includes a signal peptide at amino acids 1-20.

5.1 Antibodies

In a specific aspect, provided herein are antibodies (e.g., monoclonal antibodies, such as chimeric or humanized, for example, composite human, antibodies) which specifically bind to an extracellular domain (ECD) of human ALK (e.g., the NTR, MAM1, LDLa, MAM2 or EGF domains of human ALK-ECD). In particular embodiments, such antibodies modulate ALK expression and/or ALK activity. In particular embodiments, such antibodies bind to ALK on the surface of a cell, and are internalized.

In a specific aspect, provided herein are antigen-binding fragments of antibodies (e.g., antigen-binding fragments of monoclonal antibodies, such as chimeric or humanized, for example, composite human, antibodies) which specifically bind to an extracellular domain (ECD) of human ALK (e.g., the NTR, MAM1, LDLa, MAM2 or EGF domains of human ALK-ECD). In particular embodiments, such antigen-binding fragments modulate ALK expression and/or ALK activity. In particular embodiments, such antigen-binding fragments bind to ALK on the surface of a cell, and are internalized.

In certain embodiments, an anti-ALK antibody described herein, or an antigen-binding fragment thereof, inhibits ALK activity (e.g., ligand-dependent ALK activity and ligand-independent ALK activity) in a cell, for example, as determined by inhibition of phosphorylation of ALK, cell proliferation, and/or cell morphology. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof inhibits ALK ligand binding to ALK receptor. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof inhibits dimerization of the ALK receptor. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof induces ALK receptor degradation. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof inhibits tumor cell proliferation or tumor growth.

In certain embodiments, an anti-ALK antibody described herein, or an antigen-binding fragment thereof, increases ALK activity in a cell, for example, as determined by increase in phosphorylation of ALK, increased cell proliferation, and/or change in cell morphology. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof increases ALK ligand binding to ALK receptor. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof reduces ALK receptor degradation. In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof increases cell growth or proliferation.

In certain embodiments, an anti-ALK antibody described herein or an antigen-binding fragment thereof specifically binds to human ALK, but not mouse ALK. In certain embodiments, antibodies or antigen-binding fragments described herein can comprise sequences that do not naturally exist within the antibody germline repertoire of an animal or mammal (e.g., human) in vivo. As used herein and unless otherwise specified, the terms “about” or “approximately” mean within plus or minus 10% of a given value or range. In instances where an integer is required, the terms mean within plus or minus 10% of a given value or range, rounded either up or down to the nearest integer.

The term “antigen-binding agent” or “binding agent” or “binding protein” refers to an agent (e.g., a protein) comprising a portion (e.g., one or more binding regions such as CDRs) that binds to ALK (e.g., the ECD of ALK), and, optionally, a scaffold or framework portion (e.g., one or more scaffold or framework regions) that allows the binding portion to adopt a conformation that promotes binding of the antigen-binding agent to an ALK polypeptide, fragment, or epitope. Examples of such antigen-binding agents include antibodies, such as a human antibody, a humanized antibody, a chimeric antibody, a recombinant antibody, a single chain antibody, a diabody, a triabody, a tetrabody, a Fab fragment, a F(ab′) 2 fragment, an IgD antibody, an IgE antibody, an IgM antibody, an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody, and fragments thereof. The antigen-binding agent can comprise, for example, an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives. Such scaffolds include, but are not limited to, antibody-derived scaffolds comprising mutations introduced to, for example, stabilize the three-dimensional structure of the binding protein as well as wholly synthetic scaffolds comprising, for example, a biocompatible polymer. See, e.g., Korndorfer et al., 2003, Proteins: Structure, Function, and Bioinformatics 53(1):121-29; and Roque et al., 2004, Biotechnol. Prog. 20:639-54. In addition, peptide antibody mimetics (“PAMs”) can be used, as well as scaffolds based on antibody mimetics utilizing fibronectin components as a scaffold.

As used herein, the terms “antibody” and “immunoglobulin” and “Ig” are terms of art and can be used interchangeably herein and refer to a molecule with an antigen binding site that specifically binds an antigen.

Antibodies can include, for example, monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, such as composite human antibodies or deimmunized antibodies, murine antibodies (e.g., mouse or rat antibodies), chimeric antibodies, synthetic antibodies, and tetrameric antibodies comprising two heavy chain and two light chain molecules. In specific embodiments, antibodies can include, but are not limited to an antibody light chain monomer, an antibody heavy chain monomer, an antibody light chain dimer, an antibody heavy chain dimer, an antibody light chain/antibody heavy chain pair, an antibody with two light chain/heavy chain pairs (e.g., identical pairs), intrabodies, heteroconjugate antibodies, single domain antibodies, monovalent antibodies, bivalent antibodies, single chain antibodies or single-chain Fvs (scFv) (e.g., including monospecific, bispecific, etc.), camelized antibodies, and affybodies. Antigen-binding fragments can include antigen-binding fragments or epitope binding fragments such as, but not limited to, Fab fragments, F(ab′) fragments, F(ab′) 2 fragments, and disulfide-linked Fvs (sdFv). In certain embodiments, antibodies described herein refer to polyclonal antibody populations.

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Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA or IgY), any class, (e.g., IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 or IgA 2 ), or any subclass (e.g., IgG 2a or IgG 2b ) of immunoglobulin molecule. In certain embodiments, antibodies described herein are IgG antibodies (e.g., human IgG), or a class (e.g., human IgG 1 , IgG 2 , IgG 3 , or IgG 4 ) or subclass thereof.

In a particular embodiment, an antibody is a 4-chain antibody unit comprising two heavy (H) chain/light (L) chain pairs, wherein the amino acid sequences of the H chains are identical and the amino acid sequences of the L chains are identical. In a specific embodiment, the H and L chain comprise constant regions, for example, human constant regions. In a yet more specific embodiment, the L chain constant region of such antibodies is a kappa or lambda light chain constant region, for example, a human kappa or lambda light chain constant region. In another specific embodiment, the H chain constant region of such antibodies comprise a gamma heavy chain constant region, for example, a human gamma heavy chain constant region. In a particular embodiment, such antibodies comprise IgG constant regions, for example, human IgG constant regions.

As used herein, an “antigen” is a moiety or molecule that contains an epitope to which an antibody can specifically bind. As such, an antigen is also specifically bound by an antibody. In a particular embodiment, the antigen to which an antibody described herein binds, is human ALK, or a fragment thereof, for example, an ECD domain of human ALK.

As used herein, an “epitope” is a term in the art and refers to a localized region of an antigen to which an antibody can specifically bind. An epitope can be a linear epitope or a conformational, non-linear, or discontinuous, epitope. In the case of a polypeptide antigen, for example, an epitope can be contiguous amino acids of the polypeptide (a “linear” epitope) or an epitope can comprise amino acids from two or more non-contiguous regions of the polypeptide (a “conformational,” “non-linear” or “discontinuous” epitope). It will be appreciated by one of skill in the art that, in general, a linear epitope may or may not be dependent on secondary, tertiary, or quaternary structure. For example, in some embodiments, an antibody binds to a group of amino acids regardless of whether they are folded in a natural three dimensional protein structure. In other embodiments, an antibody requires amino acid residues making up the epitope to exhibit a particular conformation (e.g., bend, twist, turn or fold) in order to recognize and bind the epitope.

As used herein, the terms “immunospecifically binds,” “immunospecifically recognizes,” “specifically binds,” and “specifically recognizes” are analogous terms in the context of antibodies and refer to molecules that bind to an antigen/epitope as such binding is understood by one skilled in the art. For example, a molecule that specifically binds to an antigen may bind to other peptides or polypeptides, generally with lower affinity as determined by, e.g., immunoassays, surface plasmon resonance assays, for example, Biacore™, KinExA platform (Sapidyne Instruments, Boise, Id.), or other assays known in the art. In a specific embodiment, molecules that specifically bind to an antigen bind to the antigen with a K a that is at least 2 logs, 2.5 logs, 3 logs, 4 logs or greater than the K a when the molecules bind to another antigen. In another specific embodiment, molecules that specifically bind to an antigen do not cross react with other proteins. In another specific embodiment, molecules that specifically bind to an antigen do not cross react with other non-ALK proteins.

As used herein, the term “monoclonal antibody” is a well known term of art that refers to an antibody obtained from a population of homogenous or substantially homogeneous antibodies. The term “monoclonal” is not limited to any particular method for making the antibody. Generally, a population of monoclonal antibodies can be generated by cells, a population of cells, or a cell line. In specific embodiments, a “monoclonal antibody,” as used herein, is an antibody produced by a single cell or cell line wherein the antibody immunospecifically binds to an ALK epitope (e.g., an epitope of the extracellular domain of human ALK) as determined, e.g., by ELISA or other antigen-binding or competitive binding assay known in the art or in the Examples provided herein. In particular embodiments, a monoclonal antibody can be a chimeric antibody or a humanized antibody. In particular embodiments, a monoclonal antibody can be a composite human antibody. In particular embodiments, a monoclonal antibody can be a deimmunized antibody. In certain embodiments, a monoclonal antibody is a monovalent antibody or multivalent (e.g., bivalent) antibody. In particular embodiments, a monoclonal antibody is a monospecific or multispecific antibody (e.g., bispecific antibody).

As used herein, the term “polyclonal antibodies” refers to an antibody population that includes a variety of different antibodies that immunospecifically bind to the same and/or to different epitopes within an antigen or antigens.

As used herein, the terms “variable region” or “variable domain” refer to a portion of an antibody, generally, a portion of an antibody light or heavy chain, typically corresponding to about the amino-terminal 110 to 120 amino acids in a mature heavy chain and about the amino-terminal 90 to 100 amino acids in a mature light chain. Variable regions comprise complementarity determining regions (CDRs) flanked by framework regions (FRs). Generally, the spatial orientation of CDRs and FRs are as follows, in an N-terminal to C-terminal direction: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with antigen and for the specificity of the antibody for an epitope. In a specific embodiment, numbering of amino acid positions of antibodies described herein is according to the EU Index, as in Kabat et al. (1991) Sequences of Proteins of Immunological Interest , Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises murine (e.g., mouse or rat) CDRs and human framework regions (FRs). In particular embodiments, the variable region is a primate (e.g., human or non-human primate) variable region. In certain embodiments, the variable region comprises murine (e.g., mouse or rat) CDRs and primate (e.g., human or non-human primate) framework regions (FRs). As a non-limiting example, a variable region described herein is obtained from assembling two or more fragments of human sequences into a composite human sequence.

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In certain aspects, the CDRs of an antibody can be determined according to (i) the Kabat numbering system (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and, Kabat et al. (1991) Sequences of Proteins of Immunological Interest , Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242); or (ii) the Chothia numbering scheme, which will be referred to herein as the “Chothia CDRs” (see, e.g., Chothia and Lesk, 1987, J. Mol. Biol., 196:901-917; Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948; Chothia et al., 1992, J. Mol. Biol., 227:799-817; Tramontano A et al., 1990, J. Mol. Biol. 215(1):175-82; and U.S. Pat. No. 7,709,226); or (iii) the ImMunoGeneTics (IMGT) numbering system, for example, as described in Lefranc, M.-P., 1999, The Immunologist, 7:132-136 and Lefranc, M.-P. et al., 1999, Nucleic Acids Res., 27:209-212 (“IMGT CDRs”); or (iv) MacCallum et al., 1996, J. Mol. Biol., 262:732-745. See also, e.g., Martin, A., “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering , Kontermann and Dithel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001).

With respect to the Kabat numbering system, CDRs within an antibody heavy chain molecule are typically present at amino acid positions 31 to 35, which optionally can include one or two additional amino acids, following 35 (referred to in the Kabat numbering scheme as 35A and 35B) (CDR1), amino acid positions 50 to 65 (CDR2), and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3). As is well known to those of skill in the art, using the Kabat numbering system, the actual linear amino acid sequence of the antibody variable domain can contain fewer or additional amino acids due to a shortening or lengthening of a FR and/or CDR and, as such, an amino acid's Kabat number is not necessarily the same as its linear amino acid number.

In a specific embodiment, an anti-ALK antibody provided herein or an antigen-binding fragment thereof comprises CDRs of a VL region and CDRs of a VH region of any one of antibodies Ab320-Ab332 and Ab351-Ab446 described herein, and specifically binds ALK, e.g., human ALK, for example, ALK ECD, e.g. human ALK ECD. For example, provided herein are antibodies, and antigen-binding fragments thereof, that specifically bind ALK and comprise CDRs of a VL domain in Table 13, below, and CDRs of a VH domain in Table 14, below. In a specific embodiment, an anti-ALK antibody provided herein, or an antigen-binding fragment thereof, comprises a VL region and a VH region of any one of antibodies Ab320-Ab332 and Ab351-Ab446 described herein. For example, provided herein are antibodies, and antigen-binding fragments thereof, that specifically bind ALK and comprise a VL domain in Table 13, below, and VH domain in Table 14, below.

In certain embodiments, an antibody described herein comprises a VL domain as described herein, wherein the VL domain does not comprise a signal sequence. In certain embodiments, an antibody described herein comprises a VH domain as described herein, wherein the VH domain does not comprise a signal sequence. In certain embodiments, an antibody described herein comprises a VL domain and a VH domain, wherein the VL domain does not comprise a signal sequence, and wherein the VH domain does not comprise a signal sequence. In certain embodiments, an antibody described herein comprises a VL domain, wherein the VL domain comprises a signal sequence. In certain embodiments, an antibody described herein comprises a VH domain, wherein the VH domain comprises a signal sequence. In certain embodiments, an antibody described herein comprises a VL domain and a VH domain, wherein the VL domain comprises a signal sequence, and wherein the VH domain comprises a signal sequence.

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 395 and a VH region comprising SEQ ID NO: 396 (e.g., antibody Ab320). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 397 and a VH region comprising SEQ ID NO: 398 (e.g., antibody Ab321). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 399 and a VH region comprising SEQ ID NO: 400 (e.g., antibody Ab322). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 401 and a VH region comprising SEQ ID NO: 402 (e.g., antibody Ab323). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 403 and a VH region comprising SEQ ID NO: 404 (e.g., antibody Ab324). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 405 and a VH region comprising SEQ ID NO: 406 (e.g., antibody Ab325). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 407 and a VH region comprising SEQ ID NO: 408 (e.g., antibody Ab326). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 409 and a VH region comprising SEQ ID NO: 410 (e.g., antibody Ab327). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 411 and a VH region comprising SEQ ID NO: 412 (e.g., antibody Ab328). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 413 and a VH region comprising SEQ ID NO: 414 (e.g., antibody Ab329). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 415 and a VH region comprising SEQ ID NO: 416 (e.g., antibody Ab330). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 417 and a VH region comprising SEQ ID NO: 418 (e.g., antibody Ab331). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 419 and a VH region comprising SEQ ID NO: 420 (e.g., antibody Ab332).

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In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab351). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab352). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab353). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab354). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab355). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab356). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab357). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab358). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab359). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab360). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab361). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab362).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab363). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab364). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab365). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab366). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab367). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab368). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab369). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab370). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab371). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab372). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab373). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab374).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab375). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab376). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab377). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab378). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab379). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab380). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab381). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab382). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab383). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab384). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab385). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab386).

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In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab387). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab388). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab389). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab390). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab391). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab392). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab393). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab394). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab395). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab396). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab397). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab398).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab399). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab400). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab401). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab402). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab403). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab404). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab405). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab406). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab407). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab408). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab409). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab410).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab411). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab412). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab413). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab414). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab415). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab416). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab417). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab418). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab419). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab420). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab421). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab422).

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In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 429 (e.g., antibody Ab423). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab424). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab425). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab426). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab427). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab428). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab429). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab430). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab431). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab432). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab433). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab434).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 429 (e.g., antibody 435). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 430 (e.g., antibody Ab436). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 431 (e.g., antibody Ab437). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 432 (e.g., antibody Ab438). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 433 (e.g., antibody Ab439). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 434 (e.g., antibody Ab440). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 435 (e.g., antibody Ab441). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 436 (e.g., antibody Ab442). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 437 (e.g., antibody Ab443). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 438 (e.g., antibody Ab444). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 439 (e.g., antibody Ab445). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 and a VH region comprising SEQ ID NO: 440 (e.g., antibody Ab446).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 395 (e.g., the VL region of antibody Ab320). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 397 (e.g., the VL region of antibody Ab321). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 399 (e.g., the VL region of antibody Ab322). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 401 (e.g., the VL region of antibody Ab323). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 403 (e.g., the VL region of antibody Ab324). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 405 (e.g., the VL region of antibody Ab325). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 407 (e.g., the VL region of antibody Ab326). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 409 (e.g., the VL region of antibody Ab327). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 411 (e.g., the VL region of antibody Ab328). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 413 (e.g., the VL region of antibody Ab329). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 415 (e.g., the VL region of antibody Ab330). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 417 (e.g., the VL region of antibody Ab331). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 419 (e.g., the VL region of antibody Ab332).

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In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 421 (e.g., the VL region of antibody Ab351). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 422 (e.g., the VL region of antibody Ab363). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 423 (e.g., the VL region of antibody Ab375). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 424 (e.g., the VL region of antibody Ab387). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 425 (e.g., the VL region of antibody Ab399). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 426 (e.g., the VL region of antibody Ab411). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 427 (e.g., the VL region of antibody Ab423). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VL region comprising SEQ ID NO: 428 (e.g., the VL region of antibody Ab435).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 396 (e.g., the VH region of antibody Ab320). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 398 (e.g., the VH region of antibody Ab321). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO:400 (e.g., the VH region of antibody Ab322). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 402 (e.g., the VH region of antibody Ab323). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 404 (e.g., the VH region of antibody Ab324). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 406 (e.g., the VH region of antibody Ab325). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 408 (e.g., the VH region of antibody Ab326). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 410 (e.g., the VH region of antibody Ab327). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 412 (e.g., the VH region of antibody Ab328). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 414 (e.g., the VH region of antibody Ab329). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 416 (e.g., the VH region of antibody Ab330). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 418 (e.g., the VH region of antibody Ab331). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 420 (e.g., the VH region of antibody Ab332).

In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 429 (e.g., the VH region of antibody Ab351). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 430 (e.g., the VH region of antibody Ab352). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO:431 (e.g., the VH region of antibody Ab353). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 432 (e.g., the VH region of antibody Ab354). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 433 (e.g., the VH region of antibody Ab355). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 434 (e.g., the VH region of antibody Ab356). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 435 (e.g., the VH region of antibody Ab357). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 436 (e.g., the VH region of antibody Ab358). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 437 (e.g., the VH region of antibody Ab359). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 438 (e.g., the VH region of antibody Ab360). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 439 (e.g., the VH region of antibody Ab361). In a specific embodiment, an anti-ALK antibody described herein or an antigen-binding fragment thereof comprises a VH region comprising SEQ ID NO: 440 (e.g., the VH region of antibody Ab362).

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In certain embodiments, provided herein is an anti-ALK antibody or an antigen-binding fragment thereof which specifically binds to an ECD of human ALK and comprises VL CDRs (e.g., Kabat CDRs, Chothia CDRs, or IMGT CDRs) of a VL comprising the amino acid sequence as set forth in Table 13 and VH CDRs (e.g., Kabat CDRs, Chothia CDRs, or IMGT CDRs) of a VH comprising the amino acid sequence as set forth in Table 14.

In certain embodiments, provided herein is an antibody or an antigen-binding fragment thereof which specifically binds to an ECD of human ALK and comprises VL and VH CDRs of any of the anti-ALK antibodies provided herein, for example as set forth in Tables 1 and 2.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQNVGTNVA (SEQ ID NO:13);

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and

(c) a VL CDR3 comprising the amino acid sequence of QX 1 YNSYPYMX 2 T (SEQ ID NO:468), wherein X 1 is any amino acid, for example, Q or R and X 2 is any amino acid, for example, an amino acid with an aromatic side chain, e.g., Y or F. In certain embodiments, any one of X 1 and X 2 is any amino acid sequence. In specific embodiments, X 1 is a conservative substitution of Q or R, and X 2 is a conservative substitution of Y or F. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQZ 10 VZ 11 TZ 12 VA (SEQ ID NO:500), wherein Z 10 is any amino acid, for example, N or D, Z 11 is any amino acid, for example, G or S, and Z 12 is any amino acid, for example, A or N;

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14) or WASTRHT (SEQ ID NO:20) or SASNRFT (SEQ ID NO:36); and

(c) a VL CDR3 comprising the amino acid sequence of QQHYSTPRT (SEQ ID NO:21) or QQYSSYPLT (SEQ ID NO:37) or QX 1 YNSYPYMX 2 T (SEQ ID NO:468), wherein X 1 is any amino acid, for example, Q or R and X 2 is any amino acid, for example, an amino acid with an aromatic side chain, Y or F. In certain embodiments, any one of X 1 and X 2 and Z 10 -Z 12 is any amino acid sequence. In specific embodiments, X 1 is a conservative substitution of Q or R, X 2 is a conservative substitution of Y or F, Z 10 is a conservative substitution of N or D, Z 11 is a conservative substitution of G or S, and Z 12 is a conservative substitution of A or N. In specific embodiments, X 1 is a conservative substitution of Q or R, X 2 is a conservative substitution of Y or F, Z 10 is N, Z 11 is G, and Z 12 is A or N. In specific embodiments, X 1 is a conservative substitution of Q or R, X 2 is a conservative substitution of Y or F, Z 10 is D, Z 11 is S, and Z 12 is A or N. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQZ 10 VZ 11 TZ 12 VA (SEQ ID NO:500), wherein Z 10 is N or D, Z 11 is G or S, and Z 12 is A or N;

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and

(c) a VL CDR3 comprising the amino acid sequence of QQYNSYPYMYT (SEQ ID NO:15). In specific embodiments, Z 10 is N, Z 11 is G, and Z 12 is A or N. In specific embodiments, Z 10 is D, Z 11 is S, and Z 12 is A or N. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQZ 10 VZ 11 TZ 12 VA (SEQ ID NO:500), wherein Z 10 is N or D, Z 11 is G or S, and Z 12 is A or N;

(b) a VL CDR2 comprising the amino acid sequence of WASTRHT (SEQ ID NO:20); and

(c) a VL CDR3 comprising the amino acid sequence of QQHYSTPRT (SEQ ID NO:21. In specific embodiments, Z 10 is N, Z 11 is G, and Z 12 is A or N. In specific embodiments, Z 10 is D, Z 11 is S, and Z 12 is A or N. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQZ 10 VZ 11 TZ 12 VA (SEQ ID NO:500), wherein Z 10 is N or D, Z 11 is G or S, and Z 12 is A or N;

(b) a VL CDR2 comprising the amino acid sequence of SASNRFT (SEQ ID NO:36); and

(c) a VL CDR3 comprising the amino acid sequence of QQYSSYPLT (SEQ ID NO:37). In specific embodiments, Z 10 is N, Z 11 is G, and Z 12 is A or N. In specific embodiments, Z 10 is D, Z 11 is S, and Z 12 is A or N. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of KASQZ 10 VZ 11 TZ 12 VA (SEQ ID NO:500), wherein Z 10 N or D, Z 11 is G or S, and Z 12 is A or N;

(b) a VL CDR2 comprising the amino acid sequence of SASYRYS (SEQ ID NO:14); and

(c) a VL CDR3 comprising the amino acid sequence of QRYNSYPYMFT (SEQ ID NO:41). In specific embodiments, Z 10 is N, Z 11 is G, and Z 12 is A or N. In specific embodiments, Z 10 is D, Z 11 is S, and Z 12 is A or N. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

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As used herein, a “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a side chain with a similar charge. Families of amino acid residues having side chains with similar charges have been defined in the art. These families include amino acids with charged side chains (lysine, arginine, histidine, aspartic acid, glutamic acid), acidic side chains (aspartic acid, glutamic acid), basic side chains (lysine, arginine, histidine), uncharged polar side chains (glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), hydrophobic side chains (methionine, alanine, valine, leucine, isoleucine), neutral hydrophilic side chains (cysteine, serine, threonine, asparagine, glutamine), nonpolar side chains (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (threonine, valine, isoleucine) aromatic side chains (tyrosine, phenylalanine, tryptophan, histidine), and residues that influence chain orientation (glycine, proline).

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 13;

(b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:14; and

(c) a VL CDR3 comprising the amino acid sequence of SEQ ID NO:15 or 41. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 13;

(b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:14; and

(c) a VL CDR3 comprising the amino acid sequence of SEQ ID NO:15, 41, or 468. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 13, 19, 35, or 500;

(b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:14, 20, or 36; and

(c) a VL CDR3 comprising the amino acid sequence of SEQ ID NO:15, 21, 41, or 468. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK comprises:

(a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 13 or 483;

(b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:14 or 484; and

(c) a VL CDR3 comprising the amino acid sequence of SEQ ID NO:15. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises a light chain variable region (VL) comprising:

(a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO:483;

(b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:484; and

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab320, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab320 as set forth in Table 1 (SEQ ID NOS: 1, 2, and 3, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab321, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab321 as set forth in Table 1 (SEQ ID NOS: 7, 8, and 9, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab322, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab322 as set forth in Table 1 (SEQ ID NOS: 13, 14, and 15, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab323, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab323 as set forth in Table 1 (SEQ ID NOS: 19, 20, and 21, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab324, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab324 as set forth in Table 1 (SEQ ID NOS: 25, 26, and 27, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab325, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab325 as set forth in Table 1 (SEQ ID NOS: 30, 31, and 32, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

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In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab326, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab326 as set forth in Table 1 (SEQ ID NOS: 13, 14, and 15, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab327, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab327 as set forth in Table 1 (SEQ ID NOS: 35, 36, and 37, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab328, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab328 as set forth in Table 1 (SEQ ID NOS: 13, 14, and 41, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab329, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab329 as set forth in Table 1 (SEQ ID NOS: 42, 43, and 44, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab330, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab330 as set forth in Table 1 (SEQ ID NOS: 48, 49, and 50, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab331, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab331 as set forth in Table 1 (SEQ ID NOS: 48, 49, and 50, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab332, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab332 as set forth in Table 1 (SEQ ID NOS: 55, 56, and 57, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab423, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab423 as set forth in Table 1 (SEQ ID NOS: 483, 14, and 15, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, comprises the VL CDR1, VL CDR2, and VL CDR3 of Ab435, for example, the VL CDR1, VL CDR2, and VL CDR3 of Ab435 as set forth in Table 1 (SEQ ID NOS: 13, 484, and 15, respectively). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence X 3 YWMH (SEQ ID NO:469), wherein X 3 is any amino acid, for example, an amino acid with an uncharged polar side chain, e.g., N or S;

(b) a VH CDR2 comprising the amino acid sequence of YIX 4 PSSGYTKYNQKFKD (SEQ ID NO:470), wherein X 4 is any amino acid, for example N or K; and

(c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18). In certain embodiments, any one of X 3 or X 4 is any amino acid sequence. In specific embodiments, X 3 is a conservative substitution of N or S, X 4 is a conservative substitution of N or K. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of TAWMY (SEQ ID NO:51;

(b) a VH CDR2 comprising the amino acid sequence of RIKDKSNX 5 FASDYVESVRG (SEQ ID NO: 471), wherein X 5 is any amino acid, for example N or K; and

(c) a VH CDR3 comprising the amino acid sequence of SYGYA (SEQ ID NO:53). In certain embodiments, X 5 is any amino acid sequence. In specific embodiments, X 5 is a conservative substitution of N or K. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence X 3 YWMH (SEQ ID NO:469), wherein X 3 is any amino acid, for example, an amino acid with an uncharged polar side chain, e.g., N or S;

(b) a VH CDR2 comprising the amino acid sequence of Z 1 IZ 2 PZ 3 SGZ 4 TKYNZ 5 KFKZ 6 (SEQ ID NO: 498), wherein Z 1 is any amino acid, for example R or Y, Z 2 is any amino acid, for example D, K, or N, Z 3 is any amino acid, for example, N or S, Z 4 is any amino acid, for example, G or Y, Z 5 is any amino acid, for example, E or Q, and Z 6 is any amino acid, for example, S or D; and

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(c) a VH CDR3 comprising the amino acid sequence of DYYGSSZ 7 Z 8 FAY (SEQ ID NO:497), wherein Z 7 is any amino acid, for example, Y or S and Z 8 is any amino acid, for example, R or W. In certain embodiments, any one of X 3 and Z 1 -Z 8 is any amino acid sequence. In specific embodiments, X 3 is a conservative substitution of N or S, Z 1 is a conservative substitution of R or Y, Z 2 is a conservative substitution of D, K, or N, Z 3 is a conservative substitution of N or S, Z 4 is a conservative substitution of G or Y, Z 5 is a conservative substitution of E or Q, Z 6 is a conservative substitution of S or D, Z 7 is a conservative substitution of Y or S, and Z 8 is a conservative substitution of R or W. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of SYWZ 9 N (SEQ ID NO:499), wherein Z 9 is any amino acid, for example M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIYPGDGDTNYNGKFKG (SEQ ID NO: 5); and

(c) a VH CDR3 comprising the amino acid sequence of YYYGSKAY (SEQ ID NO:6) or SRGYFYGSTYDS (SEQ ID NO:29) or WYYGSYYAMDY (SEQ ID NO:33). In certain embodiments, Z 9 is any amino acid. In specific embodiments, Z 9 is a conservative substitution of M or V. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is any amino acid, for example N or S and Z 14 is any amino acid, for example M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIZ 15 PGDZ 16 DZ 17 NYNGKFKG (SEQ ID NO: 502) wherein Z 15 is any amino acid, for example, an aromatic amino acid, F or Y, Z 16 is any amino acid, for example A or G, and Z 17 is any amino acid, for example, A or T; and

(c) a VH CDR3 comprising the amino acid sequence of FSYDGAFAY (SEQ ID NO:24) or SRGYFYGSTYDS (SEQ ID NO:29) or WYYGSYYAMDY (SEQ ID NO:33). In certain embodiments, Z 13 -Z 17 are any amino acid. In specific embodiments, Z 13 is a conservative substitution of N or S, Z 14 is a conservative substitution of M or V, Z 15 is a conservative substitution of F or Y, Z 16 is a conservative substitution of A or G, and Z 17 is a conservative substitution of A or T. In specific embodiments, Z 13 is N or S, Z 14 is M or V, Z 15 is F or Y, Z 16 is A or G, and Z 17 is A or T. In specific embodiments, Z 13 is 5, Z 14 is M or V, Z 15 is F or Y, Z 16 is A or G, and Z 17 is A or T. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of SYWMH (SEQ ID NO:10);

(b) a VH CDR2 comprising the amino acid sequence of YIKPSSGYTKYZ 45 QKZ 46 Z 47 Z 48 (SEQ ID NO: 514) wherein Z 45 is N or A, Z 46 is F or L, Z 47 is K or Q, and Z 48 is D, E, or G; wherein Z 45 is any amino acid, for example, N or A, Z 46 is any amino acid, for example F or L, Z 47 is any amino acid, for example, K or Q, and Z 48 is any amino acid, for example D, E, or G; and

(c) a VH CDR3 comprising the amino acid sequence of DYYGSSSWFAY (SEQ ID NO:18). In certain embodiments, Z 45 -Z 48 are any amino acid. In specific embodiments, Z 45 is a conservative substitution of N or A, Z 46 is a conservative substitution of F or L, Z 47 is a conservative substitution of K or Q, and Z 48 is a conservative substitution of D, E or G. In specific embodiments, Z 45 is N or A, Z 46 is F or L, Z 47 is K or Q, and Z 48 is D, E, or G. In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YW 14 N (SEQ ID NO:501), wherein Z 13 is N or S and X 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIZ 15 PGDZ 16 DZ 17 NYNGKFKG (SEQ ID NO: 502) wherein Z 15 is F or Y, Z 16 is A or G, and Z 17 is A or T; and

(c) a VH CDR3 comprising the amino acid sequence of FSYDGAFAY (SEQ ID NO:24). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is N or S and Z 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIZ 15 PGDZ 16 DZ 17 NYNGKFKG (SEQ ID NO: 502) wherein Z 15 is F or Y, Z 16 is A or G, and Z 17 is A or T; and

(c) a VH CDR3 comprising the amino acid sequence of SRGYFYGSTYDS (SEQ ID NO:29). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is N or S and Z 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIZ 15 PGDZ 16 DZ 17 NYNGKFKG (SEQ ID NO: 502) wherein Z 15 is F or Y, Z 16 is A or G, and Z 17 is A or T; and

(c) a VH CDR3 comprising the amino acid sequence of WYYGSYYAMDY (SEQ ID NO:33). In a particular embodiment the antibody or an antigen-binding fragment thereof is an antibody which specifically binds to an ECD of ALK.

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In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is N or S and Z 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIFPGDADANYNGKFKG (SEQ ID NO:23); and

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is N or S and Z 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIYPGDGDTNYNGKFKG (SEQ ID NO: 5); and

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is N or S and Z 14 is M or V;

(b) a VH CDR2 comprising the amino acid sequence of QIYPGDGDTNYNGKFKG (SEQ ID NO: 5); and

In a particular embodiment, an antibody described herein, or an antigen-binding fragment thereof, which specifically binds to an ECD of ALK, comprises:

(a) a VH CDR1 comprising the amino acid sequence of Z 13 YWZ 14 N (SEQ ID NO:501), wherein Z 13 is S and Z 14 is M or V;