CN111492066A

CN111492066A - Methods of antibody targeting using transcripts and compositions derived therefrom

Info

Publication number: CN111492066A
Application number: CN201880082663.9A
Authority: CN
Inventors: Y·刘; P·郑; H-Y·周; P·张; L·杨
Original assignee: Childrens National Medical Center Inc
Current assignee: Childrens National Medical Center Inc
Priority date: 2017-10-20
Filing date: 2018-10-19
Publication date: 2020-08-04
Also published as: WO2019079762A1

Abstract

The present disclosure relates to a method of identifying and generating antibody-based cancer therapies. In some embodiments, methods are directed to identifying nucleic acid sequences encoding antibodies or antibody fragments from transcriptome or genomic DNA sequences of a sample. Another embodiment of the disclosure is a computer-implemented method of detecting the presence of a nucleic acid sequence encoding an antibody or antibody fragment in a transcriptome of a sample. The present disclosure also provides compositions comprising an antibody or antibody fragment, wherein the antibody or antibody fragment reacts with cancer tissue. The present disclosure also relates to methods of treating a subject diagnosed with or suspected of having a hyperproliferative disease, comprising administering to the subject any one or combination of the pharmaceutical compositions disclosed herein.

Description

Methods of antibody targeting using transcripts and compositions derived therefrom

Cross Reference to Related Applications

This application claims the benefit of priority from U.S. provisional patent application No. 62/575,369 filed on 2017, 10, 20, the disclosure of which is incorporated herein by reference.

Background

The generation of specific antibodies for cancer therapy is a major task involving a lengthy process of antigen identification, immunization, hybridoma production, and in most cases antibody humanization. There remains a need in the art for methods of identifying and generating antibodies for cancer therapy.

Disclosure of Invention

The present invention relates to a method for identifying and producing antibody-based cancer therapies. In some embodiments, the method of identifying a nucleic acid sequence encoding an antibody or antibody fragment from a transcriptome or genomic DNA sequence of a sample comprises the steps of:

a. screening the transcriptome or genomic DNA sequences of the sample for enriched nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences;

b. calculating a clonality score for the sample;

c. the nucleic acid sequences are divided into the first set of nucleic acid sequences and the second set of nucleic acid sequences according to a sequence alignment.

Optionally, after the classifying step, the method may further include the steps of: (d) identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand. It will be appreciated that the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences may be repeated one or more times.

In other aspects of the methods disclosed herein, an exemplary method may comprise the steps of:

e. screening the transcriptomes of a series of control subjects for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences;

f. calculating a clonality score for the sample;

g. identifying the most enriched CDR3 sequence from the sample or series of samples; and

h. obtaining a set of abundantly expressed nucleic acid sequences encoding antibody fragments from the series of control subjects, and comparing the nucleic acid sequences of the first set from the series of control subjects with the nucleic acid sequences of the first set from the subjects.

Steps (e) to (h) are carried out before, simultaneously with or after steps (a) to (c). Preferably, said step of identifying from the subject's transcriptome a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand is based on said step of comparing sets of abundantly expressed nucleic acid sequences from step (h).

The first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments. Suitable antibody fragments include: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and/or a constant region of an antibody. In other aspects of the first set of abundantly expressed nucleic acid sequences from the subject, the nucleic acid sequences encode a variable heavy chain and/or variable light chain portion of the antibody.

In other embodiments, the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for an antibody is determined, at least in part, by the following equation:

where Ci is the clone score of rearrangement i and N is the total number of rearrangements. Exemplary screening methods include performing FASTQ, MIXCR, and/or VDJtools functions on the transcriptome data.

Advantageously, prior to screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, aligning the amino acid sequences comprises performing an alignment of cloned sequences. One non-limiting example of a function that aligns cloned sequences includes performing an immunization SEQ function.

The sample used in the disclosed methods is a tissue sample from a subject having a hyperproliferative cellular disease. Samples that may be considered include, but are not limited to, cancer tissue from: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

Another embodiment of the invention is a computer-implemented method for detecting the presence of a nucleic acid sequence encoding an antibody or antibody fragment in a transcriptome of a sample, the method comprising: at least one processor and memory are included in the system.

a. Screening, by the at least one processor, the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences;

b. calculating, by the at least one processor, a clonality score;

c. separating, by the at least one processor, the nucleic acid sequences into the first set of nucleic acid sequences and the second set of nucleic acid sequences according to the clonality score.

The at least one processor may be further configured to (d) identify a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand from the first set of nucleic acid sequences after performing the step of classifying. The processor may repeat the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences one or more times.

The methods disclosed herein may further comprise the steps of:

f. calculating a clonality score corresponding to one or more of the nucleic acid sequences in each of the first and second sets of nucleic acid sequences;

g. dividing the nucleic acid sequences into the first set of nucleic acid sequences and the second set of nucleic acid sequences based on the clonality score; and

Steps (e) to (h) are carried out before, simultaneously with or after steps (a) to (c). Said step of identifying from the subject's transcriptome a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand is based on said step of comparing sets of abundantly expressed nucleic acid sequences from step (h).

In other embodiments, the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

In other embodiments, the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of variable heavy and/or variable light chain portions of the antibody.

In other embodiments, the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for the sample is determined, at least in part, by the equation:

wherein Ci is the clone score of rearrangement i and N is the total number of rearrangements; and wherein said classifying nucleic acids into said first set of nucleic acid sequences is performed if said score is 0.1 or higher.

In other embodiments, the screening the transcriptome of the series of samples for nucleic acid sequences encoding one or more antibody fragments comprises performing FASTQ, MIXCR, and vdjsols functions on the transcriptome data. Screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences. Alignment of cloned sequences includes performing an immunization SEQ function.

Samples contemplated include tissue samples from subjects with hyperproliferative cellular diseases, such as: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

In another embodiment, the first set of nucleic acids encoding antibodies or antibody fragments encodes antibodies capable of binding to one or more cells from the sample and/or subject.

The invention also features a method of editing a set of nucleic acid sequences encoding an antibody or antibody fragment from a sample, the method comprising:

(a) obtaining at least one nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of a variable strand from a transcriptome of the subject by the method of any one of claims 1 to 25; and

(b) repeating step (a) until a plurality of nucleic acid sequences encoding the antibody or antibody fragment are aggregated from the sample.

In other embodiments, a method of designing an antibody or antibody fragment capable of binding a cellular epitope from a sample comprises:

(a) obtaining at least one nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of a variable strand from a transcriptome of the subject by the method of any one of claims 1 to 25;

(b) repeating step (a) until a plurality of nucleic acid sequences encoding the antibody or antibody fragment are aggregated from the sample;

(c) cloning one or more nucleic acids in a vector or synthesizing the antibody by solid state chemical synthesis; and if the one or more nucleic acid sequences are cloned into a vector, (d) converting the vector into a host cell, and (e) allowing sufficient time for the recombinant production of the encoded antibody or antibody fragment by the host cell.

In other embodiments, a composition comprising an antibody or antibody fragment, the composition comprising at least one CDR sequence obtained by performing any one of the methods described above.

In other embodiments, an antibody library comprising at least one amino acid sequence obtained by performing any of the above methods.

In other embodiments, a non-transitory computer program product encoded on a computer-readable storage medium, the non-transitory computer program product comprising instructions for:

a. screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences;

b. calculating a clonality score corresponding to one or more of the nucleic acid sequences in each of the first and second sets of nucleic acid sequences;

c. separating the nucleic acid sequences into the first set of nucleic acid sequences and the second set of nucleic acid sequences based on the clonality score.

Optionally, the non-transitory computer program product of claim 30 further comprising instructions for: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand. Preferably, instructions are included for repeating the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences.

The inventors also consider instructions for the following steps:

The first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

The first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of variable heavy and/or variable light chain portions of the antibody.

The screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for the antibody is determined, at least in part, by the equation:

where Ci is the clone score of rearrangement i and N is the total number of rearrangements.

The screening of nucleic acid sequences encoding one or more antibody fragments in the transcriptome of a series of control subjects comprises performing FASTQ, MIXCR, and VDJtools functions on the transcriptome data.

Screening the subject's transcriptome for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences. The alignment of the cloned sequences comprises performing an immunizing SEQ function.

The sample is a tissue sample from a subject having a hyperproliferative cellular disease, such as cancer tissue selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

In other embodiments, the method of verifying antibody specificity uses immunofluorescence methods known in the art by comparing antibody binding to cancer and normal tissue.

In other embodiments, the composition of antibodies is broadly reactive against cancer tissue, but rarely against normal tissue. In a preferred embodiment, the composition of cancer specific antibodies has the amino acid sequence identified in sequence ID 1-9.

In other embodiments, the composition of antibodies has a narrow specificity for the tissue from which the antibody sequence was synthesized from the immune sequence data.

In other embodiments, the composition of individual cancer-specific antibodies of claim 45 has the amino acid sequence identified in sequence ID 10-18.

In other embodiments, the immune sequence-derived high abundance antibody sequences include CDR1, CDR2 and CDR3, as well as somatic mutated sequences. For example, the immune sequence-derived high abundance antibody sequence includes one of the sequences in sequence ID 19.

In other embodiments, the use of an antibody constructed from the sequence of claims 42 to 48 in cancer diagnosis, comprising immunohistochemical staining of tissue by immunofluorescence. The use is contemplated for cancer treatment. The disclosed uses include: antibodies and/or antibody-drug conjugates modified for optimal antibody-dependent cellular cytotoxicity or antibody-dependent phagocytosis. The antibodies are useful as components of bispecific antibodies. The antibodies are used as key components of chimeric antigen receptor T Cells (CART). Other aspects of the invention are described in the detailed description.

In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2206; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2208; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2210; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO 2216; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2218; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO 2220. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO. 2226; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO. 2228; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2230; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2236; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2238; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2240. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2246; heavy chain CDR2, said heavy chain CDR2 comprises an amino acid sequence set forth in SEQ ID NO: 2248; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO 2250; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO 2256; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO 2258; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2260. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2266; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2268; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown as SEQ ID NO: 2270; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2276; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2278; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO 2280. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO. 2286; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO 2288; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2290; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2296; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO 2298; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2300. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2306; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2308; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2310; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2316; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2318; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2320. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2326; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2328; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2330; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2336; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2338; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2340. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 2346; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 2348; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 2350; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2356; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2358; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2360. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2366; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2368; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2370; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2376; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2378; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2380. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2386; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2388; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2390; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2396; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2398; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO 2400. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence shown in SEQ ID NO: 2406; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2408; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2410; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2416; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2418; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO. 2420. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2426; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2428; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2430; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2436; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2438; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2440. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2446; a heavy chain CDR2, the heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 2448; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2450; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2456; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2458; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2460. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2466; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2468; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2470; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2476; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2478; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2480. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2486; a heavy chain CDR2, the heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2488; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2490; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2496; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2498; and a light chain CDR3, the light chain CDR3 including the amino acid sequence set forth in SEQ ID NO: 2500. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2506; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2508; a heavy chain CDR3, wherein the heavy chain CDR3 comprises an amino acid sequence shown as SEQ ID NO: 2510; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2516; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2518; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2520. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence as shown in SEQ ID NO: 2526; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence as shown in SEQ ID NO: 2528; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2530; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2536; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2538; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2540. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2546; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2548; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2550; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2556; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2558; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2560. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2566; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2568; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2570; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2576; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2578; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2580. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2586; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2588; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2590; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2596; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2598; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2600. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2606; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2608; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2610; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2616; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2618; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2620. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 2626; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2628; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2630; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2636; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2638; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2640. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2646; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2648; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2650; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2656; a light chain CDR2, the light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2658; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2660. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2666; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence shown in SEQ ID NO: 2668; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2670; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2676; a light chain CDR2, the light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2678; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2680. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2686; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2688; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2690; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2696; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2698; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2700. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2706; heavy chain CDR2, said heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 2708; heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence as shown in SEQ ID NO: 2710; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2716; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2718; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO 2720.

In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2204 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2204, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2214 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2214. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2224 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2224, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2234 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2234. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO 2244 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2244, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO 2254 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2254. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO 2264 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2264, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO 2274 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2274. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence set forth as SEQ ID NO. 2284 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO. 2284, and/or a light chain variable domain comprising the amino acid sequence set forth as SEQ ID NO. 2294 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO. 2294. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2304 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2304, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2314 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2314. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2324 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2324, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2334 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2334. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2344 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2344, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2354 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2354. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2364 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2364, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2374 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2374. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2384 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2384, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2394 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2394. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2404 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2404, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2414 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2414. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2424 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2424, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2434 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2434. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2444 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2444, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2454 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2454. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2464 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2464, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2474 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2474. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2484 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2484, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2494 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2494. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising an amino acid sequence as set forth in SEQ ID NO. 2504 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2504, and/or a light chain variable domain comprising an amino acid sequence as set forth in SEQ ID NO. 2514 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2514. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2524 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2524, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2534 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2534. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2544 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2544, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2554 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2554. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2564 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2564, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2574 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2574. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2584 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2584, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2594 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2594. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2604 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2604, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2614 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2614. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2624 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2624, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2634 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2634. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2644 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2644, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2654 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2654. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2664 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2664, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2674 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2674. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2684 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2684, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2694 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2694. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2704 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2704, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2714 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2714.

In another embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2202 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:2202, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2212 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 2212. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2222 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2222, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2232 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2232. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2242 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2242, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2252 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2252. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO 2262 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2262, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO 2272 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2272. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO. 2282 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2282, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO. 2292 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2292. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2302 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2302, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2312 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2312. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2322 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2322, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2332 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2332. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2342 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2342 and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2352 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2352. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO:2362 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2362, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO:2372 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2372. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO:2382 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2382, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO:2392 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2392. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2402 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2402, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2412 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2412. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID No. 2422 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2422, and/or a light chain comprising the amino acid sequence shown as SEQ ID No. 2432 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2432. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2442 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2442, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2452 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2452. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2462 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2462, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2472 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2472. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2482 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2482, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2492 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2492. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2502 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2502, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2512 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2512. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2522 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2522, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2532 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2532. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2542 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2542, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2552 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2552. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2562 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2562, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2572 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2572. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2582 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2582, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2592 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2592. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2602 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2602, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2612 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2612. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2622 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2622 and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2632 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2632. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2642 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2642, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2652 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2652. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2662 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2662, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2672 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2672. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2682 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2682, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2692 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2692. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2702 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2702, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2712 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2712.

In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO 2202 and the light chain comprises the amino acid sequence set forth in SEQ ID NO 2212. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO. 2222 and the light chain comprises the amino acid sequence set forth in SEQ ID NO. 2232. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2242 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2252. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2262 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2272. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO. 2282 and the light chain comprises the amino acid sequence set forth as SEQ ID NO. 2292. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO 2302 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2312. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO 2322 and the light chain comprises the amino acid sequence set forth in SEQ ID NO 2332. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2342 and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2352. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2362 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2372. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2382 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2392. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO 2402 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2412. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2422 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2432. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2442 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2452. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2462 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2472. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2482 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2492. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO 2502 and the light chain comprises the amino acid sequence set forth in SEQ ID NO 2512. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2522 and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2532. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2542 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2552. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO 2562 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2572. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO 2582 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2592. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2602 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2612. In certain embodiments, the heavy chain comprises the amino acid sequence set forth as SEQ ID NO:2622 and the light chain comprises the amino acid sequence set forth as SEQ ID NO: 2632. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2642 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2652. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2662 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2672. In certain embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO:2682 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 2692. In certain embodiments, the heavy chain comprises the amino acid sequence shown as SEQ ID NO 2702 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2712.

In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2202. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2212. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2222. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2232. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2242. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2252. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2262. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2272. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ id No. 2282. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2292. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2302. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2312. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2322. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2332. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2342. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2352. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2362. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ id No. 2372. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2382. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2392. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2402. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2412. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2422. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2432. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2442. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2452. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ id no: 2462. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2482. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2492. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2502. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2512. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2522. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2532. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2542. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2552. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2562. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ id No. 2572. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2582. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2592. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence that includes 2602. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2612. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2622. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2632. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2642. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2652. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2662. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2672. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2682. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO: 2692. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID No. 2702. In some embodiments, the composition or pharmaceutical composition is free of an amino acid sequence comprising SEQ ID NO 2712.

In one embodiment, the antibody or antigen binding fragment thereof is selected from the group consisting of Fab, Fab ', F (ab')2, Fv, domain antibodies, and single chain antibodies. In another embodiment, the antibody or antigen-binding fragment thereof is classified as an isotype selected from the group consisting of IgG, IgM, IgD, IgA, and IgE.

In another aspect, an antibody or antigen-binding fragment thereof that binds to tumor tissue competes with an antibody or antigen-binding portion thereof of any of the preceding aspects or embodiments.

In one embodiment, the antibody, or antigen-binding portion thereof, is humanized.

In one embodiment, the antibody or antigen-binding fragment thereof comprises the antibody or antigen-binding fragment thereof of any one of the aspects or embodiments herein, and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is lyophilized.

In one embodiment, the invention features an isolated nucleic acid encoding an antibody or antigen-binding fragment thereof described in any of the aspects and embodiments described herein. In one embodiment, a vector comprises the nucleic acid. In another embodiment, a host cell comprises the vector. In another embodiment, the host cell is a prokaryotic cell or a eukaryotic cell. In another embodiment, the eukaryotic cell is a protoplast cell, an animal cell, a plant cell, a fungal cell, a yeast cell, a mammalian cell, an avian cell, or an insect cell. In another embodiment, the mammalian cell is a CHO cell or a COS cell.

Drawings

FIG. 1(A-G) shows the computational framework for estimating and filtering immunoglobulin sequences based on RNAseq data from TCGA databases FIG. 1A is a schematic diagram of detection and selection of rearranged sequences from TCGA cohorts and frozen lung cancer tissues for curated expansion and experimental validation FIG. 1B is a view showing the number of aligned immunoglobulin reads (left) and clonotype counts (right) in different tumor samples FIG. 1C is a view showing CDR3 sequence clonality in different tumor types FIG. 1D is a profile showing the ratio of aligned immunoglobulin transcripts to total RNAse reads in all samples in TCGA L cohorts.A sample (TCGA-L9-A444-01A) has the highest immunoglobulin transcript and is labeled red and selected for further study.A FIG. 1E is a graph showing the score of all immunoglobulin clones from the most abundant sample (TCGA-L9-A444-01A) and is designated as red, and a selection for further study is based on the dot expression of the Japanese arrow and the point of the selected antibody sequence alignment.

FIG. 2(A and B) shows the results of immunofluorescence experiments demonstrating that recombinant antibodies reconstituted from the CDR3 sequences of L UAD patients cross-react with unrelated L USC and L0 UAD tissues FIG. 2A shows the reactivity of CDR 3-based antibodies to unrelated L USC and L UAD tissues, L3 USC (sample 427) is recognized by H8647, H2L 55, H2L, H2L, H2L 88 and H2L 99, while 2L UAD (sample 429) is recognized by H2L, H2L and H2L, FIG. 2B shows H2L and H2L combined with the UAD tissues of 2L and 2L USC, including control of mouse IgG 2L.

Fig. 3(a to D) shows the results of immunofluorescence experiments, which indicate that H2L 7 binds to various malignant tissues, but shows limited binding to normal tissues fig. 3A shows limited binding between H2L 7 and normal tissues fig. 3B shows that H2L 7 shows broad cross-reactivity to various malignant tissues, including colon adenocarcinoma, esophageal adenocarcinoma, gastric adenocarcinoma, ovarian adenocarcinoma, soft tissue giant cell tumor, liver hepatocellular carcinoma, breast infiltrating ductal carcinoma, skin squamous cell carcinoma, testicular seminoma, L UAD and L usc, fig. 3C shows a selective image depicting binding of H2L 7 to lung squamous cell carcinoma, but not to Normal Adjacent Tissues (NAT), although some non-cellular surface binding to normal parotid, normal pancreatic tissue, normal testicular tissue, normal lymph node tissue and normal spleen tissue is observed, but H2L 7 does not respond to most normal tissues fig. 3D is a fee-view of the squamous cell carcinoma binding of H2L 7 to lung and 16/16 and 4/16.

Figure 4(a to C) shows recombinant antibodies derived from L UAD (sample 463) target genomic sequences while L USC (sample 427) shows limited cross-reactivity figure 4A is a table showing selected sequences with annotation information and gene usage for further extensive analysis and experimental validation figure 4B is a view showing recombinant antibodies expressing heavy and light chains from L UAD and L USC samples by transient transfection figure 4C shows the results of immunofluorescence studies where H5L 12 derived from L UAD sample 463 recognizes 2/2L UAD tissue while H6L 13 derived from L USC sample 427 shows very specific binding to L USC tissue from which it was derived.

FIG. 5 shows an antibody library of 13 recombinant antibodies generated by random pairing of 3 heavy and 6 light chains bound to L USC and L UAD tissues, including isotype control of mouse IgG2a k as a control.

Detailed Description

The generation of specific antibodies for cancer therapy is a major task involving a lengthy process of antigen identification, immunization, hybridoma production, and in most cases antibody humanization. Unlike conventional methods, the present disclosure describes rapid large-scale identification of cancer-specific antibodies directly from human cancer tissues by de novo assembly from transcriptome and genomic sequences. The platform described herein greatly reduces the obstacles presented in the development of human anti-cancer antibodies and lays the foundation for cancer therapy using patient-derived, tumor-reactive monoclonal antibodies.

Further areas of applicability of the present disclosure will become apparent from the detailed description provided herein. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the disclosure, are intended for purposes of illustration only and are not intended to limit the scope of the disclosure.

I. Definition of

Unless defined otherwise herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by one of ordinary skill in the art. The meaning and scope of terms should be clear, but in the case of any potential ambiguity, the definitions provided herein take precedence over any dictionary or external definition. Furthermore, unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular. In the present application, the use of "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "including" as well as other forms, such as "includes" and "included," is non-limiting. Also, terms such as "element" or "component" include elements and components that comprise one unit and elements and components that comprise more than one subunit unless specifically stated otherwise.

Generally, the nomenclature and techniques described herein relating to cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization are well known and commonly employed in the art. Unless otherwise indicated, the methods and techniques disclosed herein are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. Enzymatic reactions and purification techniques were performed according to the manufacturer's instructions, as is commonly done in the art or as described herein. The nomenclature used in connection with analytical chemistry, synthetic organic chemistry, and pharmaceutical and medicinal chemistry, and the laboratory procedures and techniques described herein are those well known and commonly employed in the art. Standard techniques are used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, delivery and patient treatment.

Throughout, ranges are used as shorthand for describing each and every value that is within the range. Any value within the range can be selected as the end point value of the range. In addition, the references cited herein are incorporated by reference in their entirety. In the event that a definition in this disclosure conflicts with a definition in a cited reference, the present disclosure controls.

In order to make the present disclosure more comprehensible, selected terms are defined below.

The articles "a" and "an" are used herein to refer to one or more (i.e., to at least one) of the grammatical object of the article. For example, "an element" refers to one element or more than one element, such as a plurality of elements.

The term "including" is used herein to mean and is used interchangeably with the phrase "including but not limited to" or "including but not limited to".

The term "or" is used herein to mean and is used interchangeably with the term "and/or" unless the context clearly dictates otherwise. For example, an amino acid sequence with modified amino acids is understood to include a selection of amino acids with modified side chains, amino acids with modified backbones, and amino acids with modified side chains and modified backbones.

The term "about" as used herein means within the tolerances typical in the art. For example, "about" can be understood as about 2 standard deviations from the mean. According to certain embodiments, "about" means ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, ± 0.9%, ± 0.8%, ± 0.7%, ± 0.6%, ± 0.5%, 0.4%, 0.3%, ± 0.2%, + 0.1%, or + 0.05%. According to certain embodiments, "about" means ± 5%. When "about" is present before a series of numbers or ranges, it is understood that "about" can modify each number in the series or range.

The term "at least" preceding a plurality or series of numbers (e.g., "at least two") is to be understood to include numbers that are adjacent to the term "at least," and all subsequent numbers or integers that may be included in logic are known from the context. When a series of numbers or ranges is preceded by the word "at least," it is understood that each number in the series or range can be modified.

As used herein, "up" of "up to 10" can be understood to mean up to and include 10, i.e., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

Ranges provided herein are to be understood to include all individual integer values and all subranges within the range.

The term "polypeptide" as used herein refers to any polymer chain of amino acids. The terms "peptide" and "protein" are used interchangeably with the term polypeptide, and also refer to a polymer chain of amino acids. The term "polypeptide" encompasses natural or artificial proteins, protein fragments and polypeptide analogs of a protein sequence. The polypeptide may be monomeric or polymeric.

The term "isolated protein" or "isolated polypeptide" is a protein or polypeptide that is substantially free of other proteins of the same species, expressed by cells of a different species, or does not occur in nature due to its source or origin being unrelated to the native component with which it is associated in nature. Thus, a polypeptide that is chemically synthesized or synthesized in a cellular system different from the cell from which it is naturally derived will be "isolated" from its naturally associated components. Proteins can also be rendered substantially free of naturally associated components by isolation using protein purification techniques well known in the art. An example of an isolated polypeptide is an isolated antibody or antigen-binding portion thereof.

The term "recovering" as used herein refers to the process of making a chemical species, such as a polypeptide, substantially free of naturally associated components by separation (e.g., using protein purification techniques well known in the art).

The term "specific binding" or "specific binding" as used herein refers to the interaction of an antibody, protein or peptide with a second chemical species, meaning that the interaction is dependent on the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, antibodies recognize and bind to specific protein structures, not to general proteins. If the antibody is specific for epitope "A", the presence of a molecule comprising epitope A (or free, unlabeled A) in a reaction comprising label "A" and the antibody will reduce the amount of label A bound to the antibody.

The term "antibody" as used herein broadly refers to any immunoglobulin (Ig) molecule consisting of four polypeptide chains, two heavy (H) chains and two light (L) chains or any functional fragment, mutant, variant or derivative that retains the essential epitope binding characteristics of the Ig molecule.

In full-length antibodies, each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region comprised of three domains CH1, CH2 and CH 3. each light chain is comprised of a light chain variable region (abbreviated herein as L CVR or V L) and a light chain constant region comprised of one domain C L the VH and V L regions can be further subdivided into hypervariable regions, termed Complementarity Determining Regions (CDRs), interspersed with more conserved regions, termed Framework Regions (FRs), each VH and V L is comprised of three CDRs and four FRs arranged in the order from amino to carboxy, FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. the immunoglobulin molecule can be of any type (e.g., IgG, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG 8, IgG 6866, IgG4, IgA 27, and 2) or any subclass 2.

The term "antigen-binding portion" or "antigen-binding fragment" of an antibody as used herein (or simply "antibody portion" or "antibody fragment") refers to one or more fragments of an antibody that retain the ability to specifically bind an antigen it has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody embodiments of such antibodies can also be in bispecific, bispecific or multispecific form, i.e., specifically bind to two or more different antigens examples of binding fragments encompassed within the term "antigen-binding portion" or "antigen-binding fragment" of an antibody include (i) Fab fragments, monovalent fragments consisting of V L, VH, C L and CH1 domains, (ii) F (ab')₂Fragments which are bivalent fragments comprising two Fab fragments linked by a disulfide bond of the hinge region, (iii) Fd fragments consisting of the VH and CH1 domains, (iv) Fv fragments consisting of the V L and VH domains of a single arm of an antibody, (V) dAb fragments (Ward et al, (1989) Nature 341:544-546 and Winter et al, PCTpublication WO 90/05144A1, which are incorporated herein by reference), which comprise a single variable domain, and (vi) isolated Complementarity Determining Regions (CDRs). furthermore, although the two domains V L and VH of the Fv fragment are encoded by separate genes, they may be joined by synthetic linkers using recombinant methods, in which V L and VH regions pair to form a monovalent molecule (known as a single chain Fv scFv), see, for example, Bird et al, (1988) Science 426: 423; and Huston et al, (1988) Acad et al, (1988) Nat. 5885. antibodies also include the bivalent antibody fragment of the USA, which binds to the antigen, USA 5879. the other double antibody fragment of the USA antibodyVH and V L domains are expressed on one polypeptide chain, but the linker used is too short to pair between the two domains on the same chain, thereby forcing these domains to pair with the complementary domains of the other chain and creating two antigen binding sites (see, e.g., Holliger, P. et al, (1993) Proc. Natl. Acad. Sci. USA 90:6444-,Antibody Engineering(2001)Springer-Verlag.New York.790pp.(ISBN 3-540-41354-5)。

as used herein, the term "antibody construct" refers to a polypeptide comprising one or more antigen-binding portions of the invention linked to a linker polypeptide or immunoglobulin constant domain. Linker polypeptides comprise two or more amino acid residues joined by peptide bonds and are used to link one or more antigen binding moieties. Such linker polypeptides are known in the art (see, e.g., Holliger, P. et al, (1993) Proc. Natl. Acad. Sci. USA 90: 6444-. An immunoglobulin constant domain refers to a heavy or light chain constant domain. Human IgG heavy and light chain constant domain amino acid sequences are known in the art and are shown below.

As used herein, "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities. Isolated antibodies may be cross-reactive with other antigens from other species. In addition, the isolated antibody may be substantially free of other cellular material and/or chemicals.

The term "chimeric antibody" refers to an antibody comprising the variable region sequences of the heavy and light chains of one species and the constant region sequences of another species, e.g., a variable region antibody having murine heavy and light chains linked to human constant regions.

The term "CDR-grafted antibody" refers to an antibody that comprises heavy and light chain variable region sequences from one species, but in which the sequences of one or more CDR regions of VH and/or V L are replaced with CDR sequences from another species, e.g., an antibody having murine heavy and light chain variable regions in which one or more murine CDRs (e.g., CDR3) have been replaced with human CDR sequences.

The terms "Kabat numbering", "Kabat definitions and" Kabat labeling "are used interchangeably herein. These terms, as known in the art, refer to a system of numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody or antigen-binding portion thereof (Kabat et al, (1971) Ann. NYAcad, Sci.190: 382. 391 and Kabat, E.A. et al (1991) Sequences of Proteins of immunological Interest, Fifth Edition, U.S. department of health and public service, NIH publication No. 91-3242). For the heavy chain variable region, the hypervariable region ranged from amino acid positions 31 to 35 of CDR1, amino acid positions 50 to 65 of CDR2, and amino acid positions 95 to 102 of CDR 3. For the light chain variable region, the hypervariable region ranged from amino acid positions 24 to 34 of CDR1, amino acid positions 50 to 56 of CDR2, and amino acid positions 89 to 97 of CDR 3.

The terms "receptor" and "receptor antibody" as used herein refer to an antibody or nucleic acid sequence that provides or encodes at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or 100% of the amino acid sequence of one or more framework regions. In some embodiments, the term "receptor" refers to an antibody amino acid or nucleic acid sequence that provides or encodes a constant region. In other embodiments, the term "receptor" refers to an antibody amino acid or nucleic acid sequence that provides or encodes one or more framework regions and one or more constant regions. In a particular embodiment, the term "acceptor" refers to a human antibody amino acid or nucleic acid sequence that provides or encodes at least 80% of the amino acid sequence of one or more framework regions, preferably at least 85%, at least 90%, at least 95%, at least 98%, or 100%. According to this embodiment, the receptor may comprise at least 1, at least 2, at least 3, at least 4, at least 5, or at least 10 amino acid residues that are not (are not) present at one or more specific positions of the human antibody. The acceptor framework region and/or acceptor constant region may, for example, be derived or obtained from a germline antibody gene, a mature antibody gene, a functional antibody (e.g., an antibody well known in the art, an antibody under development, or a commercially available antibody).

The term "CDR" as used herein refers to complementarity determining regions within the variable Sequences of antibodies three CDRs in each variable region of the heavy and light chains, respectively designated CDR1, CDR2 and CDRs 3 the term "set of CDRs" as used herein refers to a set of three CDRs present in a single variable region capable of binding antigen, the defined boundaries of these CDRs have been defined differently according to different systems the system described by Kabat (Kabat et al, Sequences of Proteins of immunological Interest (National of Health, Bethesda, Md. (1987) and (1991)) not only provides a clear residue numbering system suitable for any variable region of antibodies but also provides precise residue boundaries defining three CDRs which can be designated as CDRs Chia and congeners (Chia et al, J.mol.196: 1987) and which may be found to have significant overlap with the CDRs of the CDRs 73, or even with the other CDRs found in the same CDR 27 or other CDRs 73, especially CDRs 73, or 78, respectively, although these CDRs may have significant overlap with the same amino acid residues found using the same CDR Sequences as the CDRs in the Kabat designation (see the amino acid Sequences found in the Kabat et al Sequences of the Kabat et al, No. Specification, No. Abbe found by Kabat et al, No. (see No. Abbe used herein) or No. Abbe used in the same or similar methods such as the Sect et al, the Sect et al.

The term "canonical" residues as used herein refers to residues in the CDR or framework that define the structure of a particular canonical CDR, as described by Chothia et al, J.mol.biol.196: 901-; chothia et al, J.mol.biol.227:799(1992), all of which are incorporated herein by reference. According to Chothia et al, key portions of many antibody CDRs have nearly identical peptide backbone confirmation, despite the wide variation in amino acid sequence levels. Each canonical structure specifies a set of peptide backbone torsion angles for essentially a contiguous segment of the amino acid residues that form the loop.

The terms "donor" and "donor antibody" as used herein refer to an antibody that provides one or more CDRs. In a preferred embodiment, the donor antibody is an antibody from a different species than the antibody from which the framework regions were obtained or derived. In the context of humanized antibodies, the term "donor antibody" refers to a non-human antibody that provides one or more CDRs.

The six CDRs (CDR-L1, CDR-L2 and CDR-L3 of the light chain and CDR-H1, CDR-H2 and CDR-H3 of the heavy chain) also divide the framework regions on the light and heavy chains into four subregions on each chain (FR1, FR2, FR3 and FR4), with CDR1 being located between FR1 and FR2, CDR2 being located between FR2 and FR3 and CDR3 being located between FR3 and FR 4. in the absence of designating a particular subregion as FR1, FR2, FR3 or FR4, the framework region (as otherwise indicated) represents the combined variable region within a single naturally occurring immunoglobulin chain and the combined framework regions as represented by one of the four subregions FR., as otherwise indicated, represent the four subregions of the four or more regions of the framework region as herein.

The term "fragment" as used herein is defined as a physically contiguous portion of the primary structure of a biomolecule. In some embodiments, the biomolecule is an antibody or polypeptide sequence that binds to an antigen recognized by an antibody. For an antibody or polypeptide, a fragment may be defined by a contiguous portion of the amino acid sequence of a protein, and may be at least 3 to 5 amino acids, at least 6 to 10 amino acids, at least 11 to 15 amino acids, at least 16 to 24 amino acids, at least 25 to 30 amino acids, at least 30 to 45 amino acids, and up to the full length of the protein minus a few amino acids in length. For polynucleotides, a fragment is defined by a contiguous portion of a polynucleotide nucleic acid sequence and can be at least 9 to 15 nucleotides, at least 15 to 30 nucleotides, at least 31 to 45 nucleotides, at least 46 to 74 nucleotides, at least 75 to 90 nucleotides, and at least 90 to 130 nucleotides. In some embodiments, the fragment of the biomolecule is an immunogenic fragment.

In some embodiments, the term "functional fragment" refers to any portion of a polypeptide or amino acid sequence that is of sufficient length to retain at least a portion of a biological function similar or substantially similar to the wild-type polypeptide or amino acid sequence upon which the fragment is based. If the fragment is a functional fragment of an antibody or antibody-like molecule, the fragment may be immunogenic and thus have binding affinity for one or more antigens. In some embodiments, a functional fragment of an extracellular matrix-related polypeptide is a polypeptide having 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of the polypeptides disclosed in table 1 and is of sufficient length to retain at least partial binding affinity for one or more ligands that bind to the amino acid sequences of tables 1-17 (including tables 2-16). In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in table 1, and has a length of at least about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 contiguous amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 50 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 100 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 150 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 200 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 250 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 300 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 350 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 400 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 450 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 500 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 550 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 600 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 650 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 700 amino acids. In some embodiments, the fragment is a fragment of any of the polypeptides disclosed in tables 1 to 17 and has a length of at least about 750 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 800 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 850 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 900 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 950 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 1000 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and has a length of at least about 1050 amino acids. In some embodiments, the fragment is a fragment of any of the amino acid sequences disclosed in tables 1 to 17, and is no longer than the above-described number of substitutions of the amino acids described in this paragraph. In some embodiments, a composition or pharmaceutical composition of the invention comprises any one or more of the conservative substitutions disclosed in table a.

Human heavy and light chain acceptor sequences are known in the art. In one embodiment of the invention, the human heavy and light chain acceptor sequences are selected from the sequences described in the following table.

Heavy chain receptor sequences

Light chain acceptor sequences

The term "germline antibody gene" or "gene segment" as used herein refers to immunoglobulin sequences encoded by non-lymphoid cells that have not undergone a maturation process that results in gene rearrangement and mutation to express a particular immunoglobulin. (see, e.g., Shapiro et al, Crit. Rev. Immunol.22(3):183-200 (2002); Marchalonis et al, Adv Exp Med biol.484:13-30 (2001)).

The term "hyperproliferative diseases" as used herein refers to those diseases and disorders characterized by hyperproliferation of cells or cells that exhibit dysfunction in their growth cycle, in some embodiments, the hyperproliferative diseases are dysplasia or cancer the term "hyperproliferative diseases" refers to a disease characterized by abnormal proliferation, abnormal growth, abnormal aging, abnormal quiescence or abnormal removal of cells in any organism and includes hyperplasia, neoplasia, cancer, fibroproliferative diseases (such as those involving connective tissue and other diseases characterized by fibrosis, including rheumatoid arthritis, insulin dependent diabetes, glomerulonephritis, cirrhosis and scleroderma), smooth muscle proliferative diseases (such as atherosclerosis and restenosis), chronic inflammation and epithelial cell proliferative diseases (such as psoriasis; keratosis; acne; comedogenic lesions; verrucous lesions; warts, such as verrucous warts, plantar warts, condyloma acuminatum and other diseases, such as those characterized by proliferation of epithelial cells, such as those of flat warts, cervical or thyroid, pancreatic gland, renal gland, thyroid, renal gland, thyroid, renal gland, thyroid, renal gland, adrenal gland, thyroid, renal gland, thyroid, renal gland, adrenal gland, renal gland, adrenal gland, thyroid, renal gland, adrenal gland, thyroid, renal gland, adrenal gland, thyroid, renal gland, adrenal gland, renal gland, adrenal gland, renal gland, adrenal gland, thyroid, renal gland, thyroid, adrenal gland, thyroid, adrenal gland, thyroid, renal gland, adrenal gland, thyroid, adrenal gland, thyroid, adrenal gland, renal gland, adrenal gland, thyroid, adrenal gland, renal gland, adrenal gland, renal gland, thyroid, adrenal gland, renal gland, adrenal gland, renal gland, adrenal gland, thyroid, renal gland, adrenal gland, renal gland.

The term "hyperproliferative-associated protein" as used herein refers to a protein associated with a hyperproliferative disease. In some embodiments, the hyperproliferative-associated protein is an antigen with an abnormally functional growth cycle that is expressed on a cell. In some embodiments, the cell having a dysfunctional growth cycle is cancerous. In some embodiments, the hyperproliferative-associated protein is an antigen that is expressed on hyperproliferative cells in higher amounts than normal cells. In some embodiments, the hyperproliferative-associated protein is an antigen that is expressed on hyperproliferative cells but not on normal cells. In some embodiments, the hyperproliferative cells are cells from a cancer patient sample. In some embodiments, the hyperproliferative cells are cells from cancerous tissues or solid tumors. In some embodiments, the cancer is

The term "critical" residues as used herein refers to certain residues within the variable region which have a greater effect on the binding specificity and/or affinity of an antibody, particularly a humanized antibody. Key residues include, but are not limited to, one or more of the following: residues adjacent to the CDRs, potential glycosylation sites (which can be N-or O-glycosylation sites), rare residues, residues capable of interacting with antigen, residues capable of interacting with CDRs, canonical residues, contact residues between the heavy chain variable region and the light chain variable region, residues in the vernier zone, and residues in the region where the Chothia definition of the variable heavy chain CDR1 overlaps with the Kabat definition of the first heavy chain framework.

The term "host cell" as used herein is intended to refer to a cell into which exogenous DNA has been introduced. It is understood that these terms are not intended to refer to particular subject cells, but to the progeny of such cells. Certain modifications may occur in the progeny due to mutation or environmental impact, and such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein. Preferably, the host cell comprises prokaryotic and eukaryotic cells selected from any phylum of living organisms. Eukaryotic cells include protozoan cells, fungi, plant and animal cells. Other examples include cells, but are not limited to, prokaryotic cell lines, e.coli, mammalian cell lines, CHO, HEK 293 and COS, insect cell line Sf9, and fungal cell, Saccharomyces cerevisiae.

Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipofection). enzymatic reactions and purification techniques can be performed according to the manufacturer's instructions, or as is commonly done in the art or as described herein.

The term "humanized antibody" as used herein is an antibody or variant, derivative, analog or fragment thereof which immunospecifically binds to an antigen of interest and comprises a Framework (FR) region having substantially the amino acid sequence of a human antibody and a Complementarity Determining Region (CDR) having substantially the amino acid sequence of a non-human antibody. As used herein, the term "substantially" in the context of CDRs refers to CDRs whose amino acid sequence is at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to a non-human antibody CDR. Humanized antibodies comprise substantially all of at least one and typically two variable domains (Fab, Fab ', F (ab')2, FabC, Fv), in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin (i.e., donor antibody), and all or substantially all of the framework regions are those of a human immunoglobulin consensus sequence. Preferably, the humanized antibody further comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. In some embodiments, the humanized antibody comprises both the light chain and the variable domain of the heavy chain. The antibody may also include the CH1, hinge, CH2, CH3, and CH4 regions of the heavy chain. In some embodiments, the humanized antibody comprises only a humanized light chain. In some embodiments, the humanized antibody comprises only humanized heavy chains. In particular embodiments, the humanized antibody comprises only the humanized variable domain of the light chain and/or the humanized heavy chain.

Humanized antibodies may be selected from any class of immunoglobulin including IgM, IgG, IgD, IgA, and IgE, and any isotype including, but not limited to, IgG1, IgG2, IgG3, and IgG 4. Humanized antibodies may comprise sequences from more than one type or isotype, and specific constant domains may be selected to optimize desired effector function using techniques well known in the art.

The framework regions and CDR regions of the humanized antibody do not have to correspond exactly to the parental sequences, e.g., the donor antibody CDR or consensus framework can be mutagenized by substituting, inserting, and/or deleting at least one amino acid residue such that the CDR or framework residue at that site is neither identical to the donor antibody nor the framework residue. In a preferred embodiment, however, such mutations are not widely present. Typically, at least 80%, preferably at least 85%, more preferably at least 90%, most preferably at least 95% of the humanized antibody residues will correspond to residues of the parent FR and CDR sequences. The term "consensus framework" as used herein refers to framework regions in a consensus immunoglobulin sequence. The term "consensus immunoglobulin sequence" as used herein refers to a sequence formed by the most frequently occurring amino acids (or nucleotides) in a family of related immunoglobulin sequences (see, e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987)). In the immunoglobulin family, each position in the consensus sequence is occupied by the most frequently occurring amino acid at that position in the family. If two amino acids occur equally, either can be included in the consensus sequence.

As used herein, a "vernier" region refers to a subset of framework residues that can modulate the CDR structure and fine tune the fitness to the antigen, as defined in Foote and Winter, 1992, J.Mol.biol.224:487-499, which are incorporated herein by reference. Vernier zone residues form a layer under the CDRs and may affect the structure of the CDRs and the affinity of the antibody.

The term "epitope" includes any polypeptide determinant capable of specifically binding to an antibody or antigen-binding portion thereof. In certain embodiments, epitope determinants include chemically active surface groups of a molecule (e.g., amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups), and in certain embodiments may have specific three-dimensional structural characteristics and/or specific charge characteristics. In various embodiments, the epitope can be a linear or sequential epitope of the primary structure of the antigen, i.e., a linear sequence of amino acids. Alternatively, in other embodiments, the epitope may be a conformational epitope having a particular three-dimensional shape when the antigen assumes its secondary structure. For example, a conformational epitope may include non-linear, i.e., non-sequence, amino acids of an antigen.

In a particular embodiment, the epitope is a region of an antigen bound by an antibody or antigen-binding portion thereof. In certain embodiments, an antibody or antigen-binding portion thereof is said to specifically bind to an antigen when the antibody or antigen-binding portion thereof preferentially recognizes a target antigen in a complex mixture of proteins and/or macromolecules.

The term "human antibody" as used herein is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the invention may comprise amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo), for example, in the CDRs, specifically in CDR 3. However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

The term "recombinant human antibody" as used herein is intended to include all human antibodies prepared, expressed, produced or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (further described in section II C below), antibodies isolated from recombinant combinatorial human antibody libraries (Hoogenbomom H.R., (1997) TIBTech.15: 62-70; Azzazy H., and Highsmith W.E., (2002) Clin. biochem.35: 425. 445; GaVilondo J.V.and L arick J.W. (2002) BioTechniques 29: 128. 145; Hoogenbomom H. and Chames P. (2000) Immunology Today 21: 371. 378), antibodies isolated from animals transgenic for human immunoglobulin genes (e.g.mice) (see e.g.Tannolor et al, see Taylor et al, SEQ ID NO: L; 35. 378) and antibodies derived from human immunoglobulin genes (see human antibodies) derived from human immunoglobulin genes transgenic animals transgenic human antibodies derived from human immunoglobulin genes (e.g.20, human antibodies) in vivo or from human immunoglobulin genes derived from human antibodies derived from human immunoglobulin genes (SEQ ID NO: 35, 35. Polyporas) and similar sequences found in human antibodies derived from human immunoglobulin genes (SEQ ID NO: 35, such as well as human antibodies) and human antibodies derived from human immunoglobulin genes (SEQ ID No. 35, SEQ ID No. 5, SEQ ID NO. 5, and SEQ ID No. 5, and SEQ ID NO. 5, such human antibodies derived from human antibodies, see, preferably from human antibodies derived from human.

The terms "crystal" and "crystallized" as used herein refer to an antibody or antigen-binding portion thereof that exists in a crystal form. A crystal is a form of a solid state of matter that is different from other forms such as an amorphous solid state or a liquid crystalline state. Crystals are composed of regular, repeating three-dimensional arrays of atoms, ions, molecules (e.g., proteins such as antibodies) or molecular assemblies (e.g., antigen/antibody complexes). These three-dimensional arrays are arranged according to specific mathematical relationships well known in the art. The basic unit or structural unit that repeats in the crystal is called an asymmetric unit. Repeating asymmetric units in an arrangement that conforms to a given, well-defined crystal symmetry provides a "unit cell" of the crystal. The crystal is obtained by repeating the unit cell by regular translation in all three dimensions. See, e.g., Giege, R. and Ducruix, A.Barrett, Crystallization of nucleic Acids and Proteins, a Practical Approach,2nd ea., pp.201-16, Oxford university Press, New York, New York, (1999) "

The term "polynucleotide" as used herein refers to a polymeric form of two or more nucleotides, i.e., ribonucleotides or deoxyribonucleotides or a modified form of either type of nucleotide. The term includes single-stranded and double-stranded forms of DNA, but is preferably double-stranded DNA.

The term "isolated polynucleotide" as used herein refers to a polynucleotide (e.g., of genomic, cDNA, or synthetic origin, or some combination thereof) that is "isolated polynucleotide" as a result of its origin: which is not related to all or part of a polynucleotide of which an "isolated polynucleotide" is found in nature; operably linked to a naturally unlinked polynucleotide; or not per se, as part of a larger sequence.

The term "vector" as used herein is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). After introduction into a host cell, other vectors (e.g., non-prevalent mammalian vectors) can be integrated into the genome of the host cell and thereby replicated along with the host genome. In addition, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present invention, "plasmid" and "vector" are used interchangeably as the plasmid is the most commonly used vector situation. However, the invention is intended to include other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.

The term "operably linked" refers to a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner. A control sequence "operably linked" to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences. Sequences that are "operably linked" include expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or remotely to control the gene of interest. The term "expression control sequence" as used herein refers to polynucleotide sequences necessary to effect expression and processing of coding sequences to which they are ligated. The expression control sequences include: appropriate transcription initiation, termination, promoter and enhancer sequences; effective RNA processing signals, such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequence); sequences that enhance protein stability; and sequences that enhance protein secretion when desired. The nature of such control sequences depends on the host organism; in prokaryotes, such control sequences typically include a promoter, a ribosome binding site, and a transcription termination sequence; in eukaryotes, generally, such control sequences include promoters and transcription termination sequences. The term "control sequences" is intended to include components whose presence is essential for expression and processing, and may also include other components whose presence is advantageous, such as leader sequences and fusion partner sequences. Expression vectors and host cells known in the art, including expression cassettes, vectors, recombinant host cells, and proteolytic methods for recombinant expression of recombinant proteins and proproteins from a single open reading frame, can be used to express and purify the protein constructs of the present disclosure (e.g., WO 2007/014162, which is incorporated herein by reference in its entirety).

As defined herein, "transformation" refers to any process by which foreign DNA enters a host cell. Transformation can occur under natural or artificial conditions using a variety of methods well known in the art. Transformation may rely on any known method of inserting a foreign nucleic acid sequence into a prokaryotic or eukaryotic host cell. The method is selected based on the host cell being transformed and may include, but is not limited to, viral infection, electroporation, lipofection, and particle bombardment. These "transformed" cells include stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome. They also include cells that transiently express the inserted DNA or RNA for a limited period of time.

The term "recombinant host cell (or simply" host cell "), as used herein, is intended to refer to a cell into which exogenous DNA has been introduced. It is understood that these terms are not intended to refer to particular subject cells, but to the progeny of such cells. Certain modifications may occur in the progeny due to mutation or environmental impact, and such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein. Preferably, the host cell comprises a prokaryotic and eukaryotic cell selected from any of the kingdoms of life. Preferably, eukaryotic cells include protozoan cells, fungi, plant and animal cells. Most preferably, host cells include, but are not limited to, the prokaryotic cell lines Escherichia coli, the mammalian cell lines CHO, HEK 293 and COS, the insect cell line Sf9, and the fungal cell Saccharomyces cerevisiae.

The term "inhibit" and its various grammatical forms are used to indicate to limit, block or limit the extent or degree of a certain biological event or effect.

The term "effective amount" is used herein to include an amount of an agent (e.g., a multispecific antibody) sufficient to effect treatment of a disease when administered to a patient to treat an infection in a subject (e.g., by reducing, ameliorating, or maintaining one or more symptoms of an existing disease or disease, or a complication associated therewith). The "effective amount" may vary with the drug, the mode of administration, the disease and its severity and with the medical history, age, weight, family history, genetic makeup, stage of the pathological process, type of prior or concomitant therapy (if any), and other individual characteristics of the patient to be treated. An effective amount includes an amount that causes a clinically appropriate change or stabilization of the indicator of the disease or condition. An "effective amount" refers to an amount of a compound, material, or composition described herein effective to achieve a particular biological result, such as, but not limited to, the biological results disclosed, described, or exemplified herein. Such results may include, but are not limited to, effective alleviation of symptoms associated with any of the disease states mentioned herein, as determined by any means appropriate in the art. The effective amount of the composition may depend on a number of variables including, but not limited to, species, breed, size, height, weight, age, general health of the subject, type, mode or mode of formulation or administration, type and/or severity of the particular disease being treated or the desirability of modulating the activity of the molecular pathway induced by the association of the analog with its receptor. Suitable effective amounts can be determined routinely by those skilled in the art using routine optimization techniques, with the skilled and informed judgment of the practitioner and other factors evident to those skilled in the art. An effective amount of an antibody or mutant or variant described herein can provide some or all of the biological activity as compared to the biological activity induced by the wild-type or naturally occurring polypeptide from which the antibody or mutant or variant is derived. A therapeutically effective dose of an antibody or mutant or variant described herein can provide a sustained biochemical or biological effect and/or increased resistance to degradation when placed in solution, as compared to the normal effect observed when a naturally occurring and fully processed translated protein is administered to the same subject.

An "immunoconjugate" is an antibody or multispecific antibody conjugated to one or more heterologous molecules, including but not limited to cytotoxic agents.

The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cellular function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to: a radioactive isotope; a growth inhibitor; enzymes and fragments thereof, such as nucleolytic enzymes; (ii) an antibiotic; toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.

The term "administering" as used herein means administering or applying. The term "administering" as used herein includes in vivo administration.

The term "linker" refers to a chemical moiety that links one peptide to another, e.g., an antibody to another. Linkers can also be used to link the antibody to a label or solid substrate. The linker may comprise an amino acid. The linker may be a straight or branched, saturated or unsaturated carbon chain. They may also include one or more heteroatoms within the chain.

The term "enrichment" refers to the process of identifying a transcriptome portion that is increased in number or has an increased number of nucleic acid sequences (e.g., RNA sequences). In some embodiments, the term is relative to the number of Ig-like sequences or CDR3 sequences (sequences homologous thereto) in the transcriptome of the sample. In some embodiments, the term is used to modify the number of Ig-like sequences or CDR3 sequences (sequences homologous thereto) in a transcriptome or genomic deoxyribonucleic acid (DNA) of a sample and corresponds to the abundance of nucleic acid sequences in genomic DNA or a transcriptome encoding Ig-like sequences or CDR3 sequences.

The term "pharmaceutical composition" refers to a formulation in a form that allows the biological activity of the active ingredient contained therein to be effective and does not contain other components that have unacceptable toxicity to the subject to which the composition is administered. The pharmaceutical compositions of the present invention may be administered by a variety of methods known in the art. One skilled in the art will appreciate that the route and/or mode of administration will vary depending on the desired result. In order to administer an antibody according to the invention by certain routes of administration, it may be necessary to coat or co-administer the antibody with a material that prevents its inactivation. For example, the antibody can be administered to the subject in a suitable carrier, such as a liposome or diluent. Pharmaceutically acceptable diluents include saline and buffered aqueous solutions.

By "pharmaceutically acceptable carrier" is meant an ingredient of a pharmaceutical formulation that is non-toxic to a subject, except for the active ingredient. Pharmaceutically acceptable carriers include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. In a preferred embodiment, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion).

The pharmaceutical composition according to the present invention may further comprise adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the presence of microorganisms can be ensured by the above-described sterilization procedures as well as by the inclusion of various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, and the like). It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like in the compositions. In addition, prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

The term "subject" is used throughout the specification to describe an animal that includes one or more compositions comprising one or more antibodies disclosed herein. In some embodiments, the animal is a human. For diagnosing a condition specific to a particular subject, such as a human, the term "patient" is used interchangeably. In some instances of the description of the present invention, the term "patient" will refer to a human patient suffering from a particular disease or disorder. In some embodiments, the subject may be a human suspected of having cancer or identified as at risk for having cancer. In some embodiments, the subject is suspected of having cancer or has been diagnosed with cancer. In some embodiments, the subject may be a human suspected of having cancer or identified as at risk for cancer or a hyperproliferative disease. In some embodiments, the subject may be a mammal. In some embodiments, the subject may be a non-human animal. The term "mammal" includes humans and non-humans, and includes, but is not limited to, humans, non-human primates, canines, felines, murines, bovines, equines, and porcines; as used herein, refers to an animal, such as a mammal, including a primate (e.g., a human, a non-human primate (e.g., monkey and chimpanzee)), a non-primate (e.g., a cow, pig, camel, llama, horse, goat, rabbit, sheep, hamster, guinea pig, cat, dog, rat, mouse, horse, and whale), or a bird (e.g., a duck or goose). In one embodiment, the subject is a human, such as a human treated or evaluated for a hyperproliferative disease or a human having an HIV infection that would benefit from multispecific antibodies of the invention. In some embodiments, the subject is a subject in need thereof, meaning that the subject is in need of the administered treatment.

The term "salt" refers to acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. Examples of such acids and bases are well known to those of ordinary skill in the art. Such acid addition salts are generally pharmaceutically acceptable, although salts of non-pharmaceutically acceptable acids may be used in the preparation and purification of the compounds in question. Salts include those formed from hydrochloric, hydrobromic, sulfuric, phosphoric, citric, tartaric, lactic, pyruvic, acetic, succinic, fumaric, maleic, methanesulfonic and benzenesulfonic acids. In some embodiments, a salt of a composition comprising an antibody or antibody-like molecule can be formed by reacting the free base or a salt, enantiomer or racemate thereof with one or more equivalents of an appropriate acid. In some embodiments, a pharmaceutically acceptable salt of the invention refers to a derivative or amino acid sequence that includes at least one basic group or at least one basic group. In some embodiments, pharmaceutically acceptable salts of the presently disclosed compositions include free amino, free guanidino, pyrazinyl, or pyridyl groups that form acid addition salts. In some embodiments, a pharmaceutically acceptable salt of the invention refers to a modified amino acid which is an acid addition salt of the subject compound with, for example, an inorganic acid (such as hydrochloric acid, sulfuric acid or phosphoric acid) and a suitable organic carboxylic or sulfonic acid, for example, an aliphatic mono-or dicarboxylic acid (such as trifluoroacetic acid, acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, malic acid, tartaric acid, citric acid or oxalic acid) or an amino acid (such as arginine or lysine, an aromatic carboxylic acid (such as benzoic acid, 2-phenoxy-benzoic acid, 2-acetoxybenzoic acid, salicylic acid, 4-aminosalicylic acid, an aromatic aliphatic carboxylic acid (such as mandelic acid or cinnamic acid, a heteroaromatic carboxylic acid (such as nicotinic acid or isonicotinic acid, an aliphatic sulfonic acid (such as methane-, ethane-or 2-hydroxyethanesulfonic acid or aromatic sulfonic acid))), for example benzene-, p-toluene-or naphthalene-2-sulfonic acid. When several basic groups are present, mono-or poly-acid addition salts may be formed. The reaction can be carried out in a salt-insoluble solvent or medium or in a salt-soluble solvent (e.g., water, dioxane, ethanol, tetrahydrofuran or diethyl ether or a mixture of solvents), which can be removed by vacuum or freeze-drying. The reaction may also be a metathesis process, or may be carried out on an ion exchange resin. In some embodiments, the salt may be a physiologically tolerable salt for the patient. The salts of the present invention may be found in anhydrous form or in hydrated crystalline form (i.e., completed or crystallized with one or more water molecules). In some embodiments, the composition or pharmaceutical composition comprises a crystalline form or lyophilized form of the antibody, antibody-like molecule, or salt thereof.

The term "treating" includes eliminating, substantially inhibiting, slowing or reversing the progression of a disease, disorder or disorder, substantially alleviating the clinical or aesthetic symptoms of a disorder, substantially preventing the appearance of the clinical or aesthetic symptoms of a disease, disorder or disorder, and preventing harmful or unpleasant symptoms. As used herein, the term "treating" further refers to performing one or more of the following processes: (a) reducing the severity of the disease; (b) limiting the development of symptoms characteristic of the disease being treated; (c) limiting the worsening of symptoms characteristic of the disease being treated; (d) limiting recurrence of the disease in a patient previously suffering from the disease; (e) limiting the recurrence of symptoms in previously symptomatic patients.

The term "potency" as used herein refers to the neutralizing ability, i.e. the IC of an antibody or fragment thereof₅₀Or IC₈₀。

Humanization and primatization means that an antibody can be "humanized" to reduce immunogenicity to a human recipient in the case where the trispecific fusion antibody or the three antibodies forming the trispecific fusion antibody are non-human antibodies. Methods for humanizing non-human antibodies have been described in the art. See, for example, Jones et al, Nature 321:522-525 (1986); riechmann et al, Nature 332:323-327 (1988); verhoeyen et al, Science 239:1534-1536(1988) and U.S. Pat. No.4,816,567. Typically, residues from the variable domains of non-human antibodies are "introduced" into human immunoglobulin molecules, resulting in antibodies in which some hypervariable region residues and possibly some FR residues of the human antibody are substituted by residues from analogous sites in the non-human antibody. It is important to humanize non-human antibodies while maintaining high affinity for the antigen. For this purpose, three-dimensional immunoglobulin models are generally available and are suitable for analyzing proposed humanized sequences, as compared to the parent non-human antibody. Such analysis allows the identification of residues that may be involved in antigen recognition and binding, and thus the rational design of humanized sequences that retain specificity and affinity for the antigen.

In particular embodiments, a trispecific fusion antibody is formed from any fragment or antibody sequence disclosed herein.

Similarly, a "bispecific" or "trispecific" fusion antibody or three antibodies forming a fusion can be "primatized" to reduce immunogenicity to another primate, non-human receptor, such as the rhesus receptor. "introduction" of residues from the variable domain of a donor antibody (e.g., a non-primate antibody or an antibody of a primate species other than the recipient primate) into a non-human primate recipient immunoglobulin molecule results in an antibody in which some hypervariable region residues and possibly some FR residues of the non-human primate antibody are substituted with residues from analogous sites in the donor antibody. Alternatively, primate antibodies are prepared for the desired primate species by using recipient immunoglobulins having non-primate sequences or sequences from different primate species, introducing Fc fragments and/or residues (including in particular framework region residues) from the desired primate into the recipient immunoglobulin.

"affinity maturation" refers to when one or more hypervariable region residues of an antibody can be substituted to select variants having improved biological properties relative to the parent antibody by using affinity maturation, for example using phage or yeast display. For example, the Fab region of an anti-cancer antigen can be mutated at selected several sites based on available structural information to generate all possible amino substitutions at each site. The antibody variants thus produced are displayed in monovalent form from the surface of a phage particle or yeast cell. The displayed variants are then screened for biological activity (e.g., binding affinity).

As used herein, "operably linked" in reference to a gene operably linked to a promoter refers to the linkage of two components such that expression of the gene is under the control of the promoter to which it is spatially linked. The promoter may be located 5 '(upstream) or 3' (downstream) of the gene under its control. The distance between a promoter and a gene may be about the same as the distance between the promoter and the gene it controls in the gene from which it is derived. As is known in the art, variations in this distance can be accommodated without loss of promoter function. When referring to a signal peptide operably linked to a protein, the term refers to a protein having a signal peptide incorporated as part of the protein in such a way that it can act as a signal peptide. When referring to a coding sequence that encodes a signal peptide operably linked to a coding sequence that encodes a protein, the term "coding sequence" refers to a coding sequence arranged such that translation of the coding sequence results in a protein having a portion that can be incorporated into the signal peptide in a manner that functions as a signal peptide.

As used herein, "conservative" amino acid substitutions may be as defined in table A, B or C below. Antibodies, antibody-like molecules, and derivatives, mutants, variants, and salts thereof include those amino acid sequences in which conservative substitutions have been introduced by solid state chemistry and/or recombinant modification of the nucleic acids encoding the amino acid sequences disclosed herein. In some embodiments, the compositions and pharmaceutical compositions of the invention include 1, 2, 3, 4, 5, or more conservative amino acid substitutions. Amino acids can be classified according to physical properties and contributions to secondary and tertiary protein structure. Conservative substitutions are considered in the art to be the substitution of one amino acid for another with similar properties. Exemplary conservative substitutions are shown in table a.

TABLE A-conservative substitutions I

Alternatively, conserved amino acids can be classified according to L ehninger (Biochemistry, Second Edition; Worth Publishers, inc. ny, n.y. (1975), pp.71-77), as shown in table B.

TABLE B conservative substitutions II

Uncharged-polarity

Alternatively, exemplary conservative substitutions are shown in table C.

TABLE C-conservative substitutions III

It will be appreciated that the amino acids disclosed herein are intended to include polypeptides with insertion, deletion or substitution of one or more amino acid residues, truncation or any combination, and modifications other than insertion, deletion or substitution of amino acid residues.

The term "sample" as used herein refers to a biological sample obtained or derived from a source of interest as described herein. In some embodiments, the source of interest comprises an organism, such as an animal or human. In some embodiments, the biological sample comprises a biological tissue or fluid. In some embodiments, the biological sample may be or include bone marrow, blood cells, ascites, tissue or fine needle biopsy samples, cell-containing bodily fluids, free floating nucleic acids, sputum, saliva, urine, cerebrospinal fluid, peritoneal fluid, pleural fluid, feces, lymph, gynecological fluids, swabs, vaginal swabs, oral swabs, nasal swabs, washing or lavage fluids (such as ductal or bronchoalveolar lavage fluids), aspirates, scrapings, bone marrow specimens, tissue biopsy specimens, surgical specimens, stool, other bodily fluids, secretions and/or excretions and/or cells produced therefrom, and the like. In some embodiments, the biological sample is or includes a bodily fluid. In some embodiments, the sample is a "raw sample" obtained directly from a target source by any suitable means. For example, in some embodiments, the raw biological sample is obtained by: biopsies (e.g., fine needle or tissue biopsies), surgery, collection of bodily fluids (e.g., blood, lymph, stool, etc.), and the like. In some embodiments, as is clear from the context, the term "sample" refers to a preparation obtained by processing (e.g., by removing one or more components of a primary sample and/or by adding one or more reagents thereto). For example, filtration using a semipermeable membrane. Such "processed sample" may include, for example, nucleic acids or proteins extracted from the sample, or obtained by subjecting the original sample to techniques such as amplification or reverse transcription of mRNA, separation and/or purification of certain components, etc. In some embodiments, the methods disclosed herein do not include a treated sample. In some embodiments, the methods disclosed herein comprise: obtaining a sample from water or other environmental surface, treating the sample to include a known volume, and exposing the sample to an antibody, antibody fragment, system, or composition disclosed herein.

The term "transcriptome" as used herein refers to a nucleic acid sequence that is transcribed by one or more cells in a sample. In some embodiments, the sample comprises one or more hyperproliferative cells.

As used herein, "whole blood" refers to blood obtained directly from a subject and not treated with filtration or additives prior to manipulation. In some embodiments, the whole blood may include an anticoagulant. In some embodiments, the whole blood does not contain an anticoagulant.

"sequence identity" as used herein is determined by explosion of two sequences (B12seq) using an independently executable B L AST engine program, which can be retrieved from the National Center for Biotechnology Information (NCBI) ftp site using default parameters (Tatusova and Madden, FEMS Microbiol L et., 1999,174, 247-250; which is incorporated herein by reference in its entirety.) the term "homology" is used to refer to a measured "sequence identity". in some embodiments, if one embodiment comprises a nucleic acid sequence or amino acid sequence having a percent sequence identity, the term refers to the disclosed nucleic acid sequence or amino acid sequence having homology to the disclosed sequence over its entire length.

The term "clonality score" refers to the numerical solution obtained after solving the IgH values corresponding to those nucleic acid sequences from the samples or those samples having a more extensive clonal expansion of B lymphocytes, whether or not the samples are cancerous tissue. In some embodiments, if the value is about 0.1 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.15 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.2 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.25 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.3 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.4 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.5 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.6 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.7 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.8 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.9 or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 1. or higher, the clonality score is considered to have a broader clonal expansion. In some embodiments, if the value is about 0.35 or higher, the clonality score is considered to have a broader clonal expansion.

Method of producing a composite material

The present invention relates to a method for identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome or genomic DNA sequence of a sample, said method comprising: a. screening the transcriptome or genomic DNA sequences of the sample for enriched nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences; b. calculating a clonality score for the sample; c. classifying at least one nucleic acid sequence into the first set or the second set of nucleic acid sequences based on sequence alignment between an Ig-like sequence in a sample or a nucleic acid homologous to a CDR3 (as compared to a V, J, D sequence in the sample or to the Ig-like sequence identified in a control set of samples). In some embodiments, the method further comprises the steps of: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand. In some embodiments, the method further comprises repeating the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing a plurality of CDR sequences from the sample. In some embodiments, the method further comprises identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences in the sample based on one or a combination of: the clonality score, presence or abundance of the enrichment of CDR3 or Ig-like nucleic acid sequences in the sample and the alignment of CDR sequences to CDR3 or Ig-like nucleic acid sequences in the sample. In some embodiments, the method further comprises: screening the transcriptomes of a series of control subjects for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences; calculating a clonality score for the series of control samples; identifying the most enriched CDR3 sequence from the sample or series of samples; and obtaining a set of abundantly expressed nucleic acid sequences encoding antibody fragments from the series of control subjects and comparing the first set of nucleic acid sequences from the series of control subjects with the first set of nucleic acid sequences from the subjects, wherein steps (e) to (h) are performed before, simultaneously with or after steps (a) to (c) corresponding to those subparts of the claims, and wherein the step of identifying from the subject's transcriptome a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable chain is based on the step of comparing the sets of abundantly expressed nucleic acid sequences from step (h) corresponding to those steps in the claims disclosed herein.

In some embodiments, the control sample is from a public database of transcriptomes. In some embodiments, the public database comprises transcriptomes from known cancer tissues. In some embodiments, the public database comprises transcriptomes from known cancer tissues that can be classified based on cancer type. In some embodiments, the public database comprises data from known cancer tissues from one or more cancer tissues identified herein. In some embodiments, the public database comprises transcriptomes from known cancer tissues, the public database is a TGCA database.

The present invention also relates to a computer-implemented method for detecting the presence of a nucleic acid sequence encoding an antibody or antibody fragment in a transcriptome of a sample, the method comprising: in a system comprising at least one processor and a memory; a. screening, by the at least one processor, the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences; b. calculating, by the at least one processor, a clonality score; and separating, by the at least one processor, the nucleic acid sequences into the first set of nucleic acid sequences and the second set of nucleic acid sequences according to the clonality score. In some embodiments, the method further comprises the steps of: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand.

In any of the disclosed methods, in some embodiments, the sample is a cancer tissue selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

A method of designing an antibody or antibody fragment capable of binding a cellular epitope from a sample, the method comprising: (a) obtaining at least one nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of a variable strand from a transcriptome of the subject by the method of any one of claims 1 to 25; and repeating step (a) until a plurality of nucleic acid sequences encoding the antibody or antibody fragment are aggregated from the sample; (b) cloning one or more nucleic acids in a vector or synthesizing the antibody by solid state chemical synthesis; and if the one or more nucleic acid sequences are cloned into a vector, (c) converting the vector into a host cell, and (d) allowing sufficient time for the host cell to recombinantly produce the encoded antibody or antibody fragment.

The present invention also relates to methods of treating a subject diagnosed with or suspected of having a hyperproliferative disease, comprising administering to the subject any one or combination of the pharmaceutical compositions disclosed herein. In some embodiments, the pharmaceutical composition comprises an effective amount of an antibody or antibody fragment disclosed herein or a salt thereof. In some embodiments, the hyperproliferative disease is selected from the group consisting of cancers of: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

Computer-implemented method

In some embodiments, the invention relates to a computer-implemented method of detecting, classifying, or editing one or more nucleic acid sequences encoding one or more antibodies or antibody fragments from a transcriptome of a sample. In some embodiments, the method comprises an internet-accessible device capable of performing one or more method steps, the method comprising: identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome of a subject, said identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome of a subject comprising: screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences; calculating a clonality score corresponding to one or more nucleic acid sequences in each of the first and second sets of nucleic acid sequences, and classifying the nucleic acid sequences into the first and second sets of nucleic acid sequences based on the clonality score.

In some embodiments, the invention relates to a system comprising a processor that performs a computer-implemented method of identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome of a subject, the method comprising: screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments, wherein the nucleic acid sequences encoding one or more antibody fragments comprise a first set of abundantly expressed nucleic acid sequences and a second set of rarely expressed nucleic acid sequences; calculating a clonality score corresponding to one or more nucleic acid sequences in each of the first and second sets of nucleic acid sequences, and classifying the nucleic acid sequences into the first and second sets of nucleic acid sequences based on the clonality score.

In some embodiments, the invention relates to a system comprising a processor that executes a computer-implemented method of detecting exit behavior of a user device accessing the internet, the method comprising screening a nucleic acid sequence encoding one or more antibody fragments in the transcriptome of a series of control subjects, wherein the nucleic acid sequence encoding one or more antibody fragments comprises a first set of abundantly-expressed nucleic acid sequences and a second set of rarely-expressed nucleic acid sequences, calculating a clonality score corresponding to one or more nucleic acid sequences in each of the first and second sets of nucleic acid sequences and dividing the nucleic acid sequences into the first and second sets of nucleic acid sequences based on the clonality score, and obtaining the set of abundantly-expressed nucleic acid sequences encoding antibody fragments from the series of control subjects and comparing the first set of nucleic acid sequences from the series of control subjects with the first set of nucleic acid sequences from the subjects based on the clonality score, and in some embodiments, the invention relates to a system comprising at least one processor and computer memory, the computer-readable code and/or other computer-readable storage means for storing the computer-readable code and/or computer-readable data for use in detecting the presence of the system, such as a computer-accessible computer-based on a computer-processing system, a computer-based on a computer-readable storage device, and/or computer-based on a computer-processing system, or computer-based on-readable storage-computer-based system-or computer-readable data-processing device, such as-processing system-or computer-readable data-processing device, or computer-based system-computer-readable storage device, or computer-readable data-computer-readable data-computer-readable storage device, or computer-computer.

In some embodiments, the invention relates to a computer-implemented method of identifying sequences from genomic DNA or transcriptome of nucleic acid sequences in a sample that are encoding antibodies or antibody fragments that bind cancer tissue more efficiently than antibodies or antibody fragments identified from clonal amplification of cells of the same tissue.

Exemplary embodiments include the following, and reference is made to the numbering of the claim-like language on page 71 of the specification:

1. a method of identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome or genomic DNA sequence of a sample, the method comprising:

b. calculating a clonality score for the sample;

2. The method of claim 1, further comprising the steps of: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand.

3. The method of claim 2, further comprising repeating the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences.

4. The method of claim 2, further comprising:

f. calculating a clonality score for the sample;

g. identifying the most enriched CDR3 sequence from the sample or series of samples;

h. obtaining a set of abundantly expressed nucleic acid sequences encoding antibody fragments from the series of control subjects and comparing the nucleic acid sequences of the first set from the series of control subjects to the first set of nucleic acid sequences from the subjects;

wherein steps (e) to (h) are performed before, simultaneously with or after steps (a) to (c);

and wherein said step of identifying from the subject's transcriptome a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand is based on said step of comparing sets of abundantly expressed nucleic acid sequences from step (h).

5. The method of any one of claims 1 to 5, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

6. The method of any one of claims 1 to 5, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of variable heavy and/or variable light portions of the antibody.

7. The method of any one of claims 1 to 6, wherein the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for a sample is determined at least in part by the equation:

8. The method of any one of claims 1 to 7, wherein said screening the transcriptome or genomic DNA of a series of samples for nucleic acid sequences encoding one or more antibody fragments comprises performing FASTQ, MIXCR, and VDJtools functions on the transcriptome data.

9. The method of any one of claims 1 to 8, wherein screening the transcriptome or genomic DNA of the sample for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences.

10. The method of claim 9, wherein the alignment of cloned sequences comprises performing an immunizing SEQ function.

11. The method of any one of claims 1 to 10, wherein the sample is a tissue sample from a subject having a hyperproliferative cellular disease.

12. The method of any one of claims 1 to 10, wherein the sample is a cancer tissue selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

13. A computer-implemented method for detecting the presence of a nucleic acid sequence encoding an antibody or antibody fragment in a transcriptome of a sample, the method comprising: at least one processor and memory are included in the system.

b. calculating, by the at least one processor, a clonality score;

14. The method of claim 13, further comprising the steps of: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand.

15. The method of claim 14, further comprising repeating the steps of identifying a nucleic acid sequence encoding at least one CDR of the variable chain and editing the plurality of CDR sequences.

16. The method of claim 14, further comprising:

g. dividing the nucleic acid sequences into the first set of nucleic acid sequences and the second set of nucleic acid sequences based on the clonality score;

17. The method of any one of claims 13 to 16, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

18. The method of any one of claims 13 to 17, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of variable heavy and/or variable light portions of the antibody.

19. The method of any one of claims 13 to 18, wherein the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for a sample is determined at least in part by the equation:

20. The method of any one of claims 16 to 19, wherein said screening the transcriptome of a series of samples for nucleic acid sequences encoding one or more antibody fragments comprises performing FASTQ, MIXCR, and vdjsols functions on the transcriptome data.

21. The method of any one of claims 13 to 20, wherein said screening the transcriptome of said subject for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences.

22. The method of claim 21, wherein the alignment of cloned sequences comprises performing an immunizing SEQ function.

23. The method of any one of claims 13 to 22, wherein the sample is a tissue sample from a subject having a hyperproliferative cellular disease.

24. The method of any one of claims 13 to 23, wherein the sample is a cancer tissue selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

25. The method of claim 1 or 13, wherein the first set of nucleic acids encoding antibodies or antibody fragments encodes antibodies capable of binding to one or more cells from the sample and/or subject.

26. A method of editing a set of nucleic acid sequences encoding an antibody or antibody fragment from a sample, the method comprising:

Compositions comprising antibodies

The present invention relates to one or more pharmaceutical compositions comprising an effective amount of one or more of the antibodies disclosed herein, antibody fragments disclosed herein, or salts thereof, and one or more pharmaceutically acceptable carriers.

The invention features antibodies or antigen-binding fragments thereof that bind to tumor tissue. In certain embodiments, the antibodies or antigen-binding fragments thereof of the invention are cross-reactive between tissue samples of different cancer types, but do not bind to normal tissue. In other embodiments, the antibodies or antigen binding fragments thereof of the invention selectively bind to one type of tumor tissue.

Exemplary cancers include, but are not limited to, acute lymphocytic leukemia (A LL), acute myelocytic leukemia (AM L), adrenal cancer, anal cancer, basal and squamous cell skin cancer, biliary tract cancer, bladder cancer, bone cancer, brain and spinal cord tumors, breast cancer, cervical cancer, chronic lymphocytic leukemia (C LL), chronic myelogenous leukemia (CM L), chronic myelomonocytic leukemia (CMM L) colorectal cancer, endometrial cancer, esophageal cancer, Ewing's family tumor, eye cancer, gall bladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), gestational trophoblastic disease, Hodgkin's lymphoma, Kaposi's sarcoma, kidney cancer, larynx and hypopharynx cancer, leukemia, liver cancer, lung cancer, carcinoid tumors, lymphoma, skin lymphoma, malignant mesothelioma, melanoma skin cancer, thymocyte skin cancer, multiple myeloma, myelodysplasia syndrome, nasal cavity and paranasal sinuses, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-hodgkin's lymphoma, non-small cell lung cancer, oral cancer, osteosarcoma, penile sarcoma, testicular cancer, pancreatic cancer, thyroid cancer, pancreatic cancer, uterine sarcoma, pancreatic cancer.

In some embodiments, the invention relates to a method comprising: antibody specificity was verified using immunofluorescence methods known in the art by comparing antibody binding to cancer and normal tissues.

In some embodiments, the invention relates to a composition that has broad reactivity against cancer tissue but little antibody against normal tissue.

In some embodiments, the invention relates to a composition of cancer specific antibodies having the amino acid sequence identified in sequence ID 1-9.

In some embodiments, the invention relates to a composition of antibodies having a narrow specificity for tissue in which antibody sequences are synthesized from immune sequence data.

In some embodiments, the invention relates to a composition comprising a single cancer specific antibody according to claim 45 having the amino acid sequence identified in sequence ID 10-18.

In some embodiments, the invention relates to immune sequence-derived high abundance antibody sequences including CDR1, CDR2, and CDR3, as well as somatic mutated sequences.

In some embodiments, the invention relates to an immune sequence derived sequence from the cancer assembly of claim 48 comprising one of the sequences of sequence ID 19.

In some embodiments, the invention relates to the use of antibodies constructed by the sequences of claims 42-48 in cancer diagnosis, comprising immunohistochemical staining of tissues by immunofluorescence.

In some embodiments, the antibodies are modified for optimal antibody-dependent cellular cytotoxicity or antibody-dependent phagocytosis.

In some embodiments, the antibody or fragment thereof or salt thereof is used as an antibody-drug conjugate.

In some embodiments, the antibodies are used as components of bispecific antibodies. In some embodiments, the antibodies are used as a key component of chimeric antigen receptor T Cells (CART).

One or more CDRs may be incorporated covalently or non-covalently into a molecule to make it an antigen binding protein.

An antigen binding protein can incorporate a CDR as part of a larger polypeptide chain, can covalently link a CDR to another polypeptide chain, or can non-covalently incorporate a CDR. The CDRs allow the antigen binding protein to specifically bind to a particular antigen of interest.

In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof having an antigen-binding region of any of the antibodies described in table 17.

In another embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2202 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:2202, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2212 or a sequence at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 2212. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2222 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2222, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2232 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2232. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2242 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2242, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2252 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2252. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO 2262 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2262, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO 2272 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2272. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO. 2282 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2282, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO. 2292 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2292. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2302 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2302, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2312 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2312. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2322 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2322, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2332 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2332. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2342 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2342, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2352 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2352. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO:2362 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2362, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO:2372 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2372. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence as set forth in SEQ ID NO:2382 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2382, and/or a light chain comprising the amino acid sequence as set forth in SEQ ID NO:2392 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2392. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2402 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2402, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2412 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2412. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID No. 2422 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2422, and/or a light chain comprising the amino acid sequence shown as SEQ ID No. 2432 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2432. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2442 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2442, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2452 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2452. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2462 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2462, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2472 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2472. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2482 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2482, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2492 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2492. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2502 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2502, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2512 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2512. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2522 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2522, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2532 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2532. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2542 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2542, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2552 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2552. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2562 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2562, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2572 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2572. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2582 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2582, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2592 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2592. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2602 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2602, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2612 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2612. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2622 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2622 and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2632 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2632. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2642 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2642, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2652 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2652. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2662 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2662, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2672 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2672. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2682 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2682, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2692 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2692. In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising: a heavy chain comprising the amino acid sequence shown as SEQ ID NO 2702 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2702, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO 2712 or a sequence having at least 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 2712.

In certain embodiments, the antibodies include heavy chain constant regions, such as IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant regions in certain embodiments of the invention, the antibodies or antigen-binding fragments are classified as isotypes selected from the group consisting of IgG, IgM, IgD, IgA, and IgE.

In addition, the antibody can include a light chain constant region, a kappa light chain constant region, or a lambda light chain constant region.

In certain embodiments of the invention, the antibody or antigen binding fragment is selected from the group consisting of Fab, Fab ', F (ab')2, Fv, domain antibodies, and single chain antibodies.

In other embodiments, an antibody or antigen-binding fragment thereof that binds to tumor tissue competes with an antibody or antigen-binding fragment thereof described in table 17. In particular embodiments, the competing antibody, or antigen-binding portion thereof, is an antibody, or antigen-binding portion thereof, that competes with any of the antibodies described herein. In one embodiment, the invention provides a competitive antibody that can compete with the antibody described herein.

The present invention provides a plurality of antibodies that are structurally characterized by the amino acid sequences of their variable domain regions. However, the amino acid sequence may undergo some changes while also retaining a high degree of binding to its specific target. More specifically, many amino acids in the variable domain region can be altered by conservative substitutions, and it is expected that the binding characteristics of the resulting antibody will not differ from the binding characteristics of the wild-type antibody sequence. Many amino acids in the antibody variable domain do not interact directly with the antigen or affect antigen binding and are not critical to determining antibody structure. For example, it is preferred to replace a predicted nonessential amino acid residue in any of the disclosed antibodies with another amino acid residue of the same class. Methods for identifying conservative substitutions of amino acids that do not eliminate antigen binding are well known in the art (see, for example, Brummell et al, biochem.32:1180-1187 (1993); Kobayashi et al, Protein Eng.12(10):879-884 (1999); and Burks et al, Proc. Natl. Acad. Sci. USA 94:412-417(1997), all of which are incorporated herein by reference in their entirety). Near et al explain in mol.Immunol.30:369-377,1993 how binding can be influenced or not by site-directed mutagenesis. Near et al believe that only the mutated residues have the potential to alter antigen binding. Most had a moderate or negative effect on binding affinity (Near et al, table 3) and bound to different forms of digoxin (Near et al, table 2).

Conservative modifications or functional equivalents of the disclosed peptides, polypeptides, or proteins (e.g., a hinge region or a heavy chain having a hinge region) refer to polypeptide derivatives of the peptides, polypeptides, or proteins, such as proteins having one or more point mutations, insertions, deletions, truncations, fusion proteins, or combinations thereof. Which substantially retains the activity of a parent peptide, polypeptide or protein, such as those disclosed herein. Typically, conservative modifications or functional equivalents are at least 60% (e.g., any number between 60% and 100%, including, for example, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, and 99%) identical to a parent (e.g., one of SEQ ID NOs: 1-53).

In one embodiment, a substitution that is at least 95% identical (or at least 96% identical, or at least 97% identical, or at least 98% identical, or at least 99% identical) in the heavy or light chain is a conservative amino acid substitution. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with another amino acid residue having a similar chemical property (e.g., charge or hydrophobicity) with a side chain (R group). In general, conservative amino acid substitutions do not substantially alter the functional properties of the protein. In the case where two or more amino acid sequences differ from each other by conservative substitutions, the percentage of sequence identity or similarity may be adjusted upward to correct for the conservative nature of the substitution. Methods of making such adjustments are well known to those of ordinary skill in the art. See, for example, Pearson (1994) Methods mol. biol.24: 307-. Examples of amino acid groups having side chains with similar chemical properties include: (1) aliphatic side chain: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic hydroxyl side chain: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chain: phenylalanine, tyrosine and tryptophan; (5) basic side chain: lysine, arginine and histidine; (6) acidic side chain: aspartic acid and glutamic acid; (7) the sulfur-containing side chains are cysteine and methionine.

As used herein, the percent homology between two amino acid sequences is equivalent to the percent identity between the two sequences. Given the number of gaps and the length of each gap (which need to be introduced to achieve optimal alignment of the two sequences), the percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e.,% homology-number of identical positions/total number of positions x 100). Comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the following non-limiting examples.

The algorithms of e.meyers and w.miller (comput. appl. biosci.,4:11-17(1988)), which have incorporated the a L IGN program (version 2.0), can be used to determine the percent identity between two sequences using the PAM120 weight residue table, GAP length penalty 12 and GAP penalty 4. in addition, Needleman and Wunsch (j.mol. biol.48:444-453(1970)) algorithms, which have incorporated the GAP program in the GCG software package (reviewed on www.gcg.com), can be used to determine the percent identity between two sequences using either the Blossum62 matrix or the PAM250 matrix, the

GAP weights

16, 14, 12, 10, 8, 6 or 4, the

length weights

1, 2, 3, 4,5 or 6.

Additionally or alternatively, protein sequences of the present disclosure may also be used as "query sequences" to search public databases, for example to identify related sequences, these searches may be conducted using the XB L AST program (version 2.0) of Altschul et al (1990) j.mol.biol.215:403-10. B L AST protein searches may be conducted using the XB L AST program, with a score of 50 and a word length of 3, to obtain amino acid sequences homologous to the molecules of the present invention.

Other modified forms of antibodies are contemplated herein. For example, the antibody can be linked to one of a variety of non-protein polymers, e.g., polyethylene glycol, polypropylene glycol, polyoxyalkylene, or a copolymer of polyethylene glycol and polypropylene glycol. The antibody may also be embedded in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly (methylmethacylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions. Such techniques are disclosed, for example, in the Remington's Pharmaceutical Sciences 16 th edition Oslo, A., Ed. (1980).

Variant antibodies and salts thereof are included within the scope of the invention. Variants of the sequences described herein are also included within the scope of the invention. Other variants of antibodies with improved affinity may be obtained using methods known in the art and are included within the scope of the invention. For example, amino acid substitutions may be used to obtain antibodies with further improved affinity. Alternatively, codon optimization of the nucleotide sequence can be used to increase the translation efficiency of antibody production in an expression system. Variants may include up to a certain percentage of unnatural amino acids. In some embodiments, the antibody comprises a variant amino acid sequence comprising about 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more percent of unnatural amino acids

Antibody modification

Humanization

In the case of antibodies or antigen-binding fragments thereof that are non-human antibodies, the antibodies may be "humanized" to reduce immunogenicity to human recipients. Methods for humanizing non-human antibodies have been described in the art. See, for example, Jones et al, Nature321:522-525 (1986); riechmann et al, Nature 332:323-327 (1988); verhoeyen et al, Science 239:1534-1536(1988) and U.S. Pat. No.4,816,567. Typically, residues from the variable domains of non-human antibodies are "introduced" into human immunoglobulin molecules, resulting in antibodies in which some hypervariable region residues and possibly some FR residues of the human antibody are substituted by residues from analogous sites in the non-human antibody. It is important to humanize non-human antibodies while maintaining high affinity for the antigen. For this purpose, three-dimensional immunoglobulin models are generally available and are suitable for analyzing proposed humanized sequences, as compared to the parent non-human antibody. Such analysis allows the identification of residues that may be involved in antigen recognition and binding, and thus the rational design of humanized sequences that retain specificity and affinity for the antigen.

As described above, a humanized antibody is an antibody molecule from an antibody of a non-human species that binds a desired antigen having one or more Complementarity Determining Regions (CDRs) from the non-human species and framework regions from a human immunoglobulin molecule. Known human Ig sequences have been disclosed, e.g., www.ncbi.nlm.nih.gov/entrez-/query. fcgi; www.atcc.org/phase/hdb. html; www.sciquest.com/; www.abcam.com/; www.antibodyresource.com/onlinecomp. html;

www.public.iastate.edu/.about.pedro/research_tools.html；www.mgen.uni-heidelberg.de/SD/IT/IT.html；www.whfreeman.com/immunology/CH-05/kuby05.htm；www.library.thinkquest.org/12429/Immune/Antibody.html；

www.hhmi.org/grants/lectures/1996/vlab/；www.path.cam.ac.uk/.about.mrc7/m-ikeimages.html；www.antibodyresource.com/；mcb.harvard.edu/BioLinks/Immuno-logy.html.www.immunologylink.com/；pathbox.wustl.edu/.about.hcenter/index.-html；

www.biotech.ufl.edu/.about.hcl/；www.pebio.com/pa/340913/340913.html-；

www.nal.usda.gov/awic/pubs/antibody/；www.m.ehime-u.acjp/.about.yasuhito-/Elisa.html；www.biodesign.com/table.asp；www.icnet.uk/axp/facs/davies/lin-ks.html；

www.biotech.ufl.edu/.about.fccl/protocol.html；www.isac-net.org/sites_geo.html；aximtl.imt.uni-marburg.de/.about.rek/AEP-Start.html；

baserv.uci.kun.nl/.about.jraats/linksl.html；www.recab.uni-hd.de/immuno.bme.nwu.edu/；

www.mrc-cpe.cam.ac.uk/imt-doc/pu-blic/INTRO.html；

www.ibt.unam.mx/vir/V_mice.html；imgt.cnusc.fr:8104/；

www.biochem.ucl.ac.uk/.about.martin/abs/index.html；antibody.bath.ac.uk/；

abgen.cvm.tamu.edu/lab/wwwabgen.html；www.unizh.ch/.about.honegger/AHOsem-inar/Slide01.html；www.cryst.bbk.ac.uk/.about.ubcg07s/；

www.nimr.mrc.ac.uk/CC/ccaewg/ccaewg.htm；www.path.cam.ac.uk/.about.mrc7/h-umanisation/TAHHP.html；www.ibt.unam.mx/vir/structure/stat_aim.html；

www.biosci.missouri.edu/smithgp/index.html；www.cryst.bioc.cam.ac.uk/.abo-ut.fmolina/Web-pages/Pept/spottech.html；www.jerini.de/fr roducts.htm；

www.patents.ibm.com/ibm. html, Sequences of Proteins of immunological Interest, U.S. dept. health (1983) by Kabat et al, all of which are incorporated herein by reference. Such introduced sequences may be used to reduce immunogenicity or to reduce, enhance or modify binding, affinity, turn-on rate, turn-off rate, affinity, specificity, half-life, or any other suitable characteristic known in the art.

The framework residues in the human framework regions may be substituted by corresponding residues from CDR donor antibodies to alter, preferably improve, antigen binding, these framework substitutions are identified by methods well known in the art, e.g.by modeling the interaction of the CDR and framework residues to identify framework residues important for antigen binding, and by sequence comparison to identify aberrant framework residues at specific positions (see, e.g.Queen et al, U.S. Pat. No.5,585,089; Riechmann et al, 332:323(1988), which are incorporated herein by reference in their entirety). the three-dimensional immunoglobulin model is commonly available and is familiar to the skilled person.A computer program is available which illustrates and shows the possible three-dimensional conformational structure of the selected candidate immunoglobulin sequence. examination of these display phenomena allows analysis of the possible role of residues in the function of the candidate immunoglobulin sequence, i.e.analysis of the residues affecting the ability of the candidate immunoglobulin to bind their antigen, i.e.the introduction of the residues from consensus sequences and binding to the sequences, e.e.the antibodies of WO 36419252,3685, 3685, 3699, 3685, 3670, 369, 3670, 369, 3695, 369.

Affinity maturation

One or more hypervariable region residues of an antibody can be substituted to select variants having improved biological properties relative to the parent antibody by using affinity maturation, for example using phage or yeast display. For example, the Fab region of the antibodies of the invention can be mutated at selected several sites based on available structural information to generate all possible amino substitutions at each site. The antibody variants thus produced are displayed in monovalent form from the surface of a phage particle or yeast cell. The displayed variants are then screened for biological activity (e.g., binding affinity).

Modification of Fc region

Antibodies can be modified to improve certain biological properties of the antibody, for example, to improve antibody stability, to enhance or reduce effector function (e.g., antigen-dependent cell-mediated cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC)) of the antibody, to improve or reduce internalization and/or recovery, and the like.

For example, the Fc fragment of certain antibodies (derived from human Ig4) may be substituted with human IgG1, which human IgG1 increases effector functions mediated by FcR (except FcRn). Such modifications can increase the stability of the resulting antibody by 5-fold. In another example, the IgG1 Fc fragment can be modified to improve recycling of antibodies through antibody rescue pathways.

Another type of modification involves alteration of the glycosylation pattern of the parent antibody, including deletion of one or more carbohydrate moieties found in the parent antibody, or addition of one or more carbohydrates not present in the parent antibody (by addition of one or more glycosylation sites).

Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising an antibody or antigen-binding portion thereof as exemplified herein and a pharmaceutically acceptable carrier.

The pharmaceutical compositions of the invention may comprise a "therapeutically effective amount" or a "prophylactically effective amount" of an antibody or antibody portion of the invention. A "therapeutically effective amount" is an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of an antibody or antibody portion can be determined by one of skill in the art and will vary depending on factors such as the disease state, age, sex, and weight of the individual and the ability of the antibody or antibody portion to elicit a desired response in the individual. A therapeutically effective amount is also an amount by which any toxic or deleterious effects of the antibody or antibody portion are offset by a therapeutically beneficial effect. A "prophylactically effective amount" is an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, because a prophylactic dose is used in a subject prior to or at an earlier stage of the disease, the prophylactically effective amount will be less than the therapeutically effective amount.

The antibodies or antibody portions of the invention can be incorporated into pharmaceutical compositions suitable for administration to a subject. Typically, the pharmaceutical composition comprises an antibody or antibody portion of the invention and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. The pharmaceutically acceptable carrier may also include minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives or buffers, which may extend the shelf-life or effectiveness of the antibody or antibody portion.

Various delivery systems are known and can be used to administer one or more antibodies of the invention or a combination of one or more antibodies of the invention and a useful prophylactic or therapeutic agent for methods of preventing, controlling, treating or ameliorating a disease or one or more symptoms thereof (e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing an antibody or antibody fragment), receptor-mediated endocytosis (see, e.g., Wu and Wu, j.biol. chem.262:4429-4432(1987)), nucleic acid construction of a portion of a retrovirus or other vector, and the like. Methods of administering the prophylactic or therapeutic agents of the invention include, but are not limited to, parenteral administration (e.g., intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous), epidural administration, intratumoral administration, and mucosal administration (e.g., intranasal and oral routes). In addition, pulmonary administration can be carried out, for example, by using an inhaler or nebulizer, and formulated with an aerosolizing agent. See, for example, U.S. patent nos.6,019,968, 5,985,320, 5,985,309, 5,934,272, 5,874,064, 5,855,913, 5,290,540, and 4,880,078; and PCT publication nos. WO92/19244, WO 97/32572, WO 97/44013, WO 98/31346 and WO 99/66903, all of which are incorporated herein by reference in their entirety. In one embodiment, the antibody, combination therapy, or composition of the invention is administered using the Alkermes AIR pulmonary drug delivery technology (Alkermes, inc., Cambridge, Mass.). In a particular embodiment, the prophylactic or therapeutic agent of the invention is administered intramuscularly, intravenously, intratumorally, orally, intranasally, pulmonary or subcutaneously. The prophylactic or therapeutic agent can be administered by any convenient route, e.g., by infusion or bolus injection, absorbed through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered with other bioactive agents. Systemic or local administration may be performed.

In one particular embodiment, it is desired to administer the pharmaceutical compositions of the present invention topically to an area in need of treatment, which can be accomplished by, for example, but not limited to, topical injection, by injection, or with the aid of an implant that is a porous or non-porous material that includes a membrane and a matrix, such as a salivary elastic membrane, a polymer, a fibrous matrix (e.g., TISSUE L), or a collagen matrix.

In another embodiment, the pharmaceutical composition of the invention may be delivered in the form of a Controlled or sustained Release system, in one embodiment, Controlled or sustained Release may be achieved using a pump (see L anger, supra; Sefton,1987, CRC crit. Ref. biomed. Eng.14: 20; Buchwald et al, 1980, Surgery 88: 507; Saudek et al, 1989, N.Engl. J. Med.321: 574.) in another embodiment, a polymeric material may be used to achieve Controlled or sustained Release of the therapy of the invention (see, for example, Medical Applications of Controlled Release, L anger and Wise (eds.), Pres, Boca Raca, Fla. (1974); Controlled Release Biosafety, Drugproduct, Deg, Polelabil, Polyacrylamide, Polyvinylacetate, polyethylene acrylate, polyethylene glycol.

A controlled Release system is discussed in the review of L anger (1990, Science 249:1527-1533) the Sustained Release formulation comprising one or more therapeutic agents of the present invention can be produced using any technique known to those skilled in the art, see, for example, U.S. Pat. No.4,526,938, PCT publication No. WO 91/05548, PCT publication No. WO 96/20698, Ning et al 1996, "Integrated Radiology of Human Colon Cancer Xenorograft Using a Sustanated-Release Gel," Radiology & Oncology 39:179-189, Song et al 1995, "organism medical 34. under Targeting of L-cloning-circulation Emulsions," PDAjournal of biological Science & Technology 50:372, control et al 1997 "see, biological samples for example # 94.22 and" biological filtration of biological samples ". 24," these references ".

In a particular embodiment, where the composition of the invention is a nucleic acid encoding a prophylactic or therapeutic agent, the nucleic acid may be administered in vivo to facilitate expression of the prophylactic or therapeutic agent encoded therein by constructing the nucleic acid as part of a suitable nucleic acid expression vector and administering it to bring the nucleic acid into the cell (e.g., by using a retroviral vector) (e.g., see U.S. Pat. No.4,980,286), or by direct injection or using microprojectile bombardment (e.g., gene gun; Biolistic, Dupont), or coated with a lipid or cell surface receptor or transfection agent, or by use in conjunction with a homeobox-shaped peptide known to enter the nucleus (e.g., see Joliot et al, 1991, Proc. Natl. Acad. Sci. USA 88: 4-1868). Alternatively, the nucleic acid may be introduced into the cell and incorporated into the host cell DNA for expression by homologous recombination.

The pharmaceutical compositions of the present invention are formulated to be compatible with their intended route of administration. Examples of routes of administration include, but are not limited to, parenteral, e.g., intravenous, intradermal, subcutaneous, oral, intranasal (e.g., inhalation), transdermal (e.g., topical), transmucosal, and rectal administration. In a particular embodiment, the composition is formulated into a pharmaceutical composition suitable for intravenous, subcutaneous, intramuscular, oral, intranasal or topical administration to a human according to conventional procedures. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. If necessary, the composition may also include a solubilizing agent and a local anesthetic (e.g., lignin) to reduce pain at the injection site.

If the compositions of the present invention are to be administered topically, the compositions may be formulated as ointments, creams, transdermal patches, lotions, gels, shampoos, sprays, aerosols, solutions, emulsions, or other forms well known to those skilled in the art. See, for example, the summary of pharmaceutical sciences and pharmaceutical dosage forms of Remington, 19 th edition, Mack pub. Co., Easton, Pa. (1995). For non-sprayable topical dosage forms, viscous to semi-solid or solid forms containing a carrier or one or more excipients compatible with topical application and having a dynamic viscosity preferably greater than water are typically used. Suitable formulations include, but are not limited to, solutions, suspensions, emulsions, creams, ointments, powders, ini's, salves, and the like, which may be sterilized or mixed or admixed with adjuvants (e.g., preservatives, stabilizers, wetting agents, buffers, or salts) to affect various properties, such as osmotic pressure, if desired. Other suitable topical dosage forms include sprayable aerosols wherein the active ingredient, preferably in combination with a solid or liquid inert carrier, is preferably packaged in admixture with a pressurized volatile substance (e.g., a gaseous propellant such as freon) or in squeeze bottles. Humectants can also be added to pharmaceutical compositions and dosage forms, if desired. Examples of other such feedstocks are known in the art.

If the method of the invention comprises intranasal administration of the composition, the composition may be formulated in the form of an aerosol, spray, mist or drops. In particular, the prophylactic or therapeutic agents for use according to the present invention may be conveniently delivered in aerosol form from pressurized packs or a nebulizer, with the use of a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges (e.g., of gelatin) for use in an inhaler may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

If the methods of the present invention include oral administration, the compositions may be formulated orally in the form of tablets, capsules, cachets, gel caps, solutions, suspensions, and the like. Tablets or capsules may be prepared by conventional means together with pharmaceutically acceptable excipients such as binding agents (e.g. pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropylmethyl cellulose), fillers (e.g. lactose, microcrystalline cellulose or dibasic calcium phosphate), lubricants (e.g. magnesium stearate, talc or silicon dioxide), disintegrants (e.g. potato starch or sodium starch glycolate) or wetting agents (e.g. sodium lauryl sulfate). The tablets may be coated by methods known in the art. Liquid preparations for oral administration may take the form of, but are not limited to, solutions, syrups or suspensions, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared in conventional manner together with pharmaceutically acceptable additives such as suspending agents (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats), emulsifying agents (for example, lecithin or acacia), non-aqueous vehicles (for example, almond oil, oily esters, ethanol or fractionated vegetable oils) and preservatives (for example, methyl or propyl p-hydroxybenzoates or sorbic acid). The formulations may also suitably contain buffer salts, flavouring agents, colouring agents and sweetening agents. Formulations for oral administration may be suitably formulated for sustained, controlled or sustained release of the prophylactic or therapeutic agent.

The methods of the invention may include pulmonary administration, for example by use of an inhaler or nebulizer, with concomitant formulation with an aerosol. See, for example, U.S. patent nos.6,019,968, 5,985,320, 5,985,309, 5,934,272, 5,874,064, 5,855,913, 5,290,540, and 4,880,078; and PCT publication nos. WO92/19244, WO 97/32572, WO 97/44013, WO 98/31346 and WO 99/66903, all of which are incorporated herein by reference in their entirety. In a particular embodiment, the antibody, combination therapy and/or composition of the invention is administered using the Alkermes AIR pulmonary drug delivery technology (Alkermes, inc., Cambridge, Mass.).

The methods of the invention may include administration of compositions formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion). Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

The methods of the invention may further comprise administering a composition formulated as a depot formulation. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compositions may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins or as sparingly soluble derivatives (e.g., as a sparingly soluble salt).

The methods of the invention comprise the administration of a composition configured as a mesoscopic or saline dosage form. Pharmaceutically acceptable salts include salts with anions and with cations, for example, anions derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, and the like, for example, cations derived from sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

Typically, the components of the composition are provided separately or mixed together in unit dosage form, e.g., as a dry lyophilized powder or as an anhydrous concentrate in a closed container, e.g., in an ampoule or sachet, to indicate the amount of active agent. Where the mode of administration is infusion, the composition can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the mode of administration is by injection, an ampoule of sterile water for injection or saline may be provided so that the ingredients may be mixed prior to administration.

In particular, the invention also provides packaging one or more prophylactic or therapeutic agents or pharmaceutical compositions of the invention in a hermetically sealed container, such as an ampoule or sachet, which indicates the amount of the agent. In one embodiment, one or more prophylactic or therapeutic agents or pharmaceutical compositions of the invention are provided in a sealed container as a dry sterile lyophilized powder or anhydrous concentrate and can be reconstituted (e.g., with water or saline) to an appropriate concentration for administration to a subject. Preferably, one or more prophylactic or therapeutic agents or pharmaceutical compositions of the invention are provided in a unit dose of at least 5mg, more preferably at least 10mg, at least 15mg, at least 25mg, at least 35mg, at least 45mg, at least 50mg, at least 75mg or at least 100mg in a closed container in the form of a dry sterile lyophilized powder. The lyophilized prophylactic or therapeutic agent or pharmaceutical composition of the present invention should be stored between 2 ℃ and 8 ℃. The prophylactic or therapeutic agent of the invention or the pharmaceutical composition of the invention should be added to its original container and should be administered within 1 week of reconstitution, preferably within 5 days, 72 hours, 48 hours, 24 hours, 12 hours, 6 hours, 5 hours, 3 hours or 1 hour thereafter. In an alternative embodiment, one or more prophylactic or therapeutic agents or pharmaceutical compositions of the invention are provided in liquid form in a sealed container to indicate the amount and concentration of the agent. Preferably, the liquid dosage form of the administration composition is provided in a sealed container of at least 0.25mg/ml, more preferably in a sealed container of at least 0.5mg/ml, at least 1mg/ml, at least 2.5mg/ml, at least 5mg/ml, at least 8mg/ml, at least 10mg/ml, at least 15mg/ml, at least 25mg/ml, at least 50mg/ml, at least 75mg/ml or at least 100 mg/ml. The liquid dosage form should be stored in its original container at between 2 ℃ and 8 ℃.

Preferably, the antibody or antibody portion of the invention will be prepared as an injection containing 0.1-250mg/ml of the antibody, preferably the injection solution may consist of a liquid or lyophilized formulation in a flint or amber vial, ampoule or pre-filled syringe, the buffer may be L-histidine (1-50mM), most preferably 5-10mM, pH 5.0 to 7.0 (most preferably pH 6.0), other suitable buffers including but not limited to sodium succinate, sodium citrate, sodium phosphate or potassium phosphate, sodium chloride may be at a concentration of 0-300mM (most preferably 150mM for a liquid formulation) to improve the toxicity of the solution, for lyophilized formulations, cryoprotectants may be included, primarily 0-10% sucrose (most preferably 0.5-1.0%) for cryoprotectants, other suitable cryoprotectants including trehalose and lactose for lyophilized formulations may include bulking agents, primarily 1-10% mannitol (most preferably 2-4%) for a liquid and stabilizing agent may be used with a liquid and stabilizing agent, and for use as a pharmaceutical composition for parenteral administration with a hyaluronic acid addition of a therapeutic agent, including as a polysorbate 0.10% increasing agent, preferably for parenteral administration of a pharmaceutical composition, e.g. polysorbate 20010% of a pharmaceutical composition, which may also include as a polysorbate-sorbitol, a parenteral adjuvant, preferably a topical adjuvant, for parenteral injection, and/or a pharmaceutical formulation for parenteral injection having an additional bioavailability of an additional suitable for use of a pharmaceutical composition including for increasing the therapeutic agent, including for parenteral protein, e.g. a pharmaceutical composition including a polysorbate, e.g. a topical formulation for parenteral injection having an additional bioavailability of a therapeutic effect, including a topical formulation including a polysorbate, e.g. No. 5-10% sorbitol, No. 5-10, No. 5-10% sorbitol, No. 5-10, No.5 for parenteral injection.

The compositions of the present invention may be in a variety of dosage forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The preferred dosage form depends on the intended mode of administration and therapeutic application. Typical preferred compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for passive immunization of humans with other antibodies. Preferred modes of administration are parenteral (e.g. intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the antibody is administered by intravenous infusion or injection in another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection

Therapeutic compositions must generally be sterile and stable under the conditions of manufacture and storage. The compositions may be formulated as solutions, microemulsions, dispersions, liposomes or other ordered structures suitable for high drug concentrations. Sterile injectable solutions can be prepared by incorporating the active compound (i.e., the antibody or antibody portion) in the required amount in an appropriate solvent with one or more of the ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile lyophilized powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and spray drying which yields a powder of the active ingredient and any additional desired ingredient from a previously sterile-filtered solution thereof. Proper fluidity of the solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, such as monostearate salts and gelatin.

Although the preferred route/mode of administration for many therapeutic applications is subcutaneous injection, intravenous injection or infusion, the antibodies and antibody portions of the invention can be administered by a variety of methods known in the art. One skilled in the art will appreciate that the route and/or mode of administration will vary depending on the desired result. In certain embodiments, the active compounds can be prepared with carriers that protect the compound from rapid release, such as controlled release formulations, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Many methods of preparing such formulations have been patented or are generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, J.R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

In certain embodiments, the antibodies or antibody portions of the invention may be administered orally, e.g., with an inert diluent or an absorbable edible carrier. The compound (and other ingredients, if desired) can also be encapsulated in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the diet of a subject. For oral therapeutic administration, the compounds may be combined with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, antiageing drugs, suspensions, syrups, wafers, and the like. In order to administer a compound of the invention by means other than parenteral administration, it may be desirable to include the compound in, or co-administer it with, a material that prevents its inactivation.

In other embodiments, the antibodies or antibody moieties of the invention may be conjugated to a polymer-based substance such that the polymer-based substance can impart to the antibodies or antibody moieties of the invention sufficient size to benefit from the antibodies or antibody moieties of the invention with enhanced permeability and retention effects (EPR effects) (see also PCT publication No. wo2006/042146a2 and U.S. patent publication nos.2004/0028687a1, 2009/0285757a1 and 2011/0217363a1, and U.S. patent No.7,695,719), all of which are incorporated herein by reference in their entirety.

In certain embodiments, the antibody or fragment thereof is linked to a carrier known in the art to increase half-life. Such carriers include, but are not limited to, Fc domains, polyethylene glycols, and dextrans. Such vectors are described in the following patents: for example, U.S. patent serial No.09/428,082 and published PCT application No. wo 99/25044, both of which are incorporated herein by reference.

In a particular embodiment, a nucleic acid sequence comprising a nucleotide sequence encoding an antibody of the invention or another prophylactic or therapeutic agent of the invention is administered by gene therapy to treat, prevent, control or ameliorate a disease or one or more symptoms thereof. Gene therapy refers to treatment by administering an expressed or expressible nucleic acid to a subject. In this embodiment of the invention, the nucleic acid produces the antibody of the invention encoded thereby or a prophylactic or therapeutic agent that mediates a prophylactic or therapeutic effect.

General reviews on Gene therapy methods are available in the art, see Goldspire et al, 1993, Clinical Pharmacy 12:488-505, Wu and Wu, 1991, Biotherapy 3:87-95, Tolstoshev 1993, Ann.Rev.Pharmacol.Toxicol.32:573-596, Mulligan Science 260:926-932(1993), and Morgan and Anderson, 1993, Ann.Rev.biochem.62:191-217, May 1993, TIBTECH 11(5): 155-215. well known methods in the field of recombinant DNA technology, as described in Aurea et al (documents), Inc. molecular Biology, Willian & William & 1990, the methods described in detail in U.S. Stodserves et al (handbook et al, USA), the methods of treatment are incorporated by the teachings of Golddserv et al, Inc., handbook et al, (1990, 19852, Inc., Prekusank.A.52).

Antibody production

The antibodies of the invention can be produced by any of a variety of techniques known in the art. For example, expression from a host cell is by standard techniques, wherein expression vectors encoding the heavy and light chains are transfected into the host cell. The term "transfection" of various forms is intended to cover the usually used to introduce exogenous DNA into prokaryotic or eukaryotic host cells in a variety of techniques, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection. Although it is possible to express the antibodies of the invention in prokaryotic or eukaryotic host cells, expression of the antibodies in eukaryotic cells is preferred, and most preferred in mammalian host cells, because such eukaryotic cells (particularly mammalian cells) are more likely than prokaryotic cells to assemble and secrete properly folded and immunologically active antibodies.

Exemplary nucleic acids of the invention are shown in table 17.

Exemplary mammalian host cells for expressing recombinant antibodies of the invention include Chinese hamster ovary (CHO cells) (including DHFR-CHO cells such as Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with DHFR selectable markers (e.g., as described in R.J. Kaufman and P.A. Sharp (1982) mol. biol.159: 601-621), NS0 myeloma cells, COS cells, and SP2 cells.

The host cell may also be used to produce functional antibody fragments, such as Fab fragments or scFv molecules. It will be appreciated that variations of the above process are within the scope of the invention. For example, it may be desirable to transfect a host cell with DNA encoding a functional fragment of the light and/or heavy chain of an antibody of the invention. Recombinant DNA techniques can also be used to remove some or all of the DNA encoding one or both of the light or heavy chains that are not necessary for binding to the antigen of interest. Molecules expressed by such truncated DNA molecules are also included in the antibodies of the invention. Alternatively, bifunctional antibodies may be produced in which one heavy and one light chain is an antibody of the invention, and the other heavy and light chains are antibodies specific for an antigen other than the antigen of interest by cross-linking the antibody of the invention to a second antibody by standard chemical cross-linking methods.

In a preferred system for recombinant expression of the antibodies of the invention, or antigen-binding portions thereof, recombinant expression vectors encoding the antibody heavy and light chains are introduced into DHFR-CHO cells by calcium phosphate-mediated transfection.

Another embodiment of the invention provides a glycosylated antibody or an antigen binding portion thereof, wherein the antibody or antigen binding portion thereof comprises one or more carbohydrate residues. The production of nascent proteins in vivo may undergo further processing, known as post-translational modification. Specifically, sugar (glycosyl) residues can be added enzymatically, a process known as glycosylation. The resulting protein with covalently attached oligosaccharide side chains is called a glycosylated protein or glycoprotein. Antibodies are glycoproteins that have one or more carbohydrate residues in the Fc domain as well as in the variable domain. Carbohydrate residues in the Fc domain have a significant effect on the effector function of the Fc domain with minimal effect on antigen binding or antibody half-life (r.jefferis, biotechnol. prog.21(2005), pp.11-16). In contrast, glycosylation of the variable domain may have an effect on the antigen binding activity of the antibody. Glycosylation in the variable domain may have a negative effect on antibody binding affinity, possibly due to steric hindrance (Co, M.S. et al, mol.Immunol. (1993)30: 1361. sup. 1367), or lead to an increased affinity for the antigen (Wallick, S.C. et al, exp.Med. (1988)168: 1099. sup. 1109; Wright, A. et al, EMBO J. (1991)10: 2717. sup. 2723).

One aspect of the invention relates to the generation of glycosylation site mutants, wherein the O-or N-linked glycosylation sites of an antibody, or antigen-binding portion thereof, have been mutated. Such mutants can be generated by one skilled in the art using standard well-known techniques. Glycosylation site mutants that retain biological activity but have increased or decreased binding activity are another object of the invention.

In yet another embodiment, the glycosylation of an antibody or antigen binding portion of the invention is modified. For example, non-glycosylated antibodies (i.e., antibodies lacking glycosylation) can be prepared. Glycosylation can be altered, for example, to increase the affinity of an antibody for an antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made that result in the elimination of one or more variable region glycosylation sites, thereby eliminating glycosylation at that site. This glycosylation can increase the affinity of the antibody for the antigen. The process is further described in detail in PCT publication No. WO2003016466a2 and U.S. patent nos.5,714,350 and 6,350,861, all of which are incorporated herein by reference in their entirety.

Additionally or alternatively, modified antibodies of the invention with altered glycosylation patterns can be prepared, such as sub-fucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNAc structure.

Protein glycosylation depends on the amino acid sequence of the protein of interest and the host cell in which the protein is expressed. Different organisms can produce different glycosylases (e.g., glycosyltransferases and glycosidases) and have different substrates (nucleotide sugars) available. Due to these factors, the glycosylation pattern of a protein and the composition of glycosyl residues may vary, depending on the host system in which a particular protein is expressed. Glycosyl residues useful in the present invention can include, but are not limited to, glucose, galactose, mannose, fucose, n-acetylglucosamine, and sialic acid. Preferably, the glycosylated antibody or antigen binding portion thereof comprises glycosyl residues such that the glycosylation pattern is human.

It is known to the person skilled in the art that different glycosylation of proteins may lead to different protein properties. For example, a therapeutic protein that is glycosylated using the yeast endogenous pathway produced in a microbial host, such as yeast, may have reduced efficacy compared to the same protein expressed in a mammalian cell, such as a CHO cell line. Such glycoproteins may also be immunogenic in humans and have a reduced half-life in vivo following administration. Specific receptors in humans and other animals recognize specific glycosyl residues and facilitate rapid clearance of proteins from the bloodstream. Other adverse effects may include changes in protein folding, solubility, sensitivity to proteases, trafficking, transport, compartmentalization, secretion, recognition by other proteins or factors, antigenicity, or allergenicity. Thus, a practitioner may prefer a therapeutic protein having a particular glycosylation composition and pattern, for example, a glycosylation composition and pattern that is the same as or at least similar to that produced in human cells or species-specific cells of the intended subject animal.

Expression of a glycosylated protein different from the host cell may be achieved by genetically modifying the host cell to express the heterologous glycosylase. Using techniques known in the art, practitioners can generate antibodies or antigen-binding portions thereof that exhibit glycosylation of human proteins. For example, yeast strains have been genetically modified to express non-naturally occurring glycosylases such that glycosylated proteins (glycoproteins) produced in these yeast strains exhibit the same protein glycosylation as animal cells, particularly as human cells (U.S. patent nos. 20040018590 and 20020137134, and PCT publication No. WO2005100584 a 2).

In addition to antibodies or antigen-binding portions thereof, the invention also relates to anti-idiotypic (anti-Id) antibodies specific for such antibodies or antigen-binding portions thereof of the invention. An anti-Id antibody is an antibody that recognizes a unique determinant that is normally associated with the antigen binding region of another antibody. anti-Id can be prepared by immunizing an animal with an antibody or antigen-binding portion thereof or a CDR-containing region thereof. The immunized animal will recognize and respond to the idiotypic determinants of the immunizing antibody and produce an anti-Id antibody. The anti-Id antibody may also be used as an "immunogen" to induce an immune response in another animal, thereby producing a so-called anti-Id antibody.

Furthermore, it will be appreciated by those skilled in the art that a library of host cells genetically engineered to express various glycosylation enzymes can be used to express a protein of interest such that the member host cells of the library produce the protein of interest with a variable glycosylation pattern. The practitioner can then select and isolate a protein of interest having a particular novel glycosylation pattern-preferably, a protein having a particular selected novel glycosylation pattern exhibits improved or altered biological properties.

It will be apparent to those skilled in the art that other suitable modifications and variations of the method of the invention described herein are apparent and may be made using suitable equivalents without departing from the scope of the invention or the embodiments disclosed herein. Having now described the invention in detail, it will be more clearly understood by reference to the following examples, which are included merely for purposes of illustration and are not intended to limit the invention.

Any and all journal articles, patent applications, issued patents, and publications disclosed herein

Or other cited references, are incorporated by reference in their respective entireties.

Examples of the invention

Example 1 computational framework for estimation and filtering of immunoglobulin sequences based on RNAseq data from the TCGA database.

The present invention describes a computational framework developed and successfully tested antibody discovery by mining large numbers of Ig CDR3 sequences in samples based on solid tumor RNA sequencing in 1945 in the TCGA databases of glioblastoma multiforme (GBM), lower gliomas (L GG), lung adenocarcinoma (L UAD), squamous cell carcinoma of the lung (L USC), pancreatic adenocarcinoma (PAAD), and cutaneous melanoma (SKCM). surprisingly, synthetic antibodies based on selected high abundance CDR3 sequences from patients L UAD bind to multiple lung cancer samples and are cross-reactive with other types of cancer but importantly have very little cross-reaction with normal tissue.

Recent studies by us and others have shown that the TCGA RNAseq database captures the characteristics of the T cell pool within the tumor (3). However, similar analysis has not been performed on antibody libraries. To fill this major gap, a platform was developed that not only describes the status of antibody responses, but also identifies specific sequences that may be useful in cancer therapy and diagnosis, as shown in fig. 1A.

First, reads are aligned to the B cell receptor reference V, D, J and C genes, and then gene signature sequences (called clone sequences) are extracted from the aligned reads, as shown in fig. 1B, identifying a large number of Ig transcripts in all six cancer types analyzed, wherein the L USC and L UAD transcriptomes contain significantly more Ig transcriptomes, next, clonotypes are assembled for the clone sequences (based on the CDR3 regions) after the core clonotypes are constructed, a mapping is performed that processes the delayed records in the previous step in order to rescue quantitative information from the low-mass reads, clustering is performed after the clonotypes are assembled by the original assembler and mapper, connecting the low-abundance clonotypes to the high-abundance, significantly similar clonotypes, as shown in fig. 1B, although the BCR clonotypes of L USC and L UAD transcriptomes are more abundant, a large number of clonotypes are found in all six cancer types, and thus, the number of millions of aligned heavy chain reads (igeq) in the heavy chain reads are used as a measure of the number of heavy chain assembled Ig transcripts per million of millions of igpm in all six cancer types.

The analysis of B cell clonality reveals substantial differences in B cell clonality between different patients between different tumor populations (fig. 1C), thus providing valuable guidance for identifying patient samples with highly amplified BCR clones, the final step is to align the cloned sequences with reference VD, J, and C genes to reconstruct the alignment of the cloned sequences and add information to them, such as amino acid sequences and related variable (V) and linking (J) genes, considering the main abundances observed in BCR density and clonality in TCGA L UAD populations, the selection of immunoglobulin transcripts relative to the abundance of total eq for the first sample ranking of the igr clones for verification of the abundance of the first sample for the igr clone abundance using the IgH sequence alignment chart for verification of the IgH sequence score in ige 3, the CDR score chart for the first sample of IgH 369 igg gene alignment chart for verification of the CDR sequence score using the IgH 75, CDR 75, and CDR 75 h score chart for the test results of the CDR sequence alignment chart, the results of the IgH 369 and CDR 75 h.

Co-enrichment of the H and L chain CDR3 suggests that they are likely to pair, as suggested by parallel studies in T cells (4) however, it is not possible to determine pairing by frequency alone (5). despite this, since the goal is to recognize cancer-reactive antibodies rather than to describe endogenous antibody responses, 3H chain and 6L chain were randomly combined in pairs and tested to determine whether immunoglobulins were expressed from these pairs.these antibodies were linked to mouse IgG2a heavy and k light chains to facilitate future studies of human tissues that may have endogenous human IgG.As shown in FIG. 1G, of the 18 combinations, 13 combinations produced significant levels of antibody in transient transfections and therefore provided a valuable tool for assessing antibody specificity.

Tables 1-6 show the top 100 heavy chains and the top 100 light chains selected from the top three patients in the TCGA database.

Tables 7-16 show the first 50 heavy chains and the first 50 light chains selected from each tumor sample of the Adaptive Biotechnologies database.

Example 2 antibody repertoire of 13 recombinant antibodies generated by random pairing of 3 heavy and 6 light chains bound to L USCs and L UAD tissues.

To determine whether de novo assembled antibodies bound to cancer tissue, these 13 antibodies were paired and immunofluorescent with frozen tissue of L USC and L UAD. the assembled antibodies were tested using goat anti-mouse second step reagents as shown in fig. 5, the antibody library showed superior binding to L USC and L UAD. when the antibodies were tested alone, 9/13 showed clear binding to L UAD. of these, 6 also reacted with L USC (fig. 2A). H2L 7 was selected for further testing because it has a unique staining pattern and has broad cross reactivity with all five lung cancer samples, including two L UAD tumor tissues and 3L USC tumor tissues in frozen sections.

Example 3H 2L 7 binds to a variety of malignant tissues, but has limited binding to normal tissues.

H2L 7 is characterized by its binding to normal and different types of cancer tissue after frozen tissue microarray examination of normal tissue, H2L 7 was unresponsive to most of the normal tissues tested (fig. 3A) although some binding to intracellular components or extracellular matrix was found in pancreas, spleen, parotid gland and testis, no definitive cell surface staining was found for any normal tissue in contrast, H2L 7 was broadly reactive to the 9/11 types of cancer tissues tested, including colon adenocarcinoma, esophageal adenocarcinoma, gastric adenocarcinoma, ovarian adenocarcinoma, soft tissue cytoma, liver hepatocellular carcinoma, testicular seminoma, lung adenocarcinoma and lung squamous carcinoma (fig. 3B) in addition, H2L 7 was tested against L USC TMA containing paired malignant and normal adjacent tissues the staining results showed that H2L 7 binds strongly to 100% of L USC but only to 25% of adjacent benign tissues (fig. 3C and 3D).

Example 4 recombinant antibodies derived from the target genomic sequence of L UAD (sample 463) whereas L USC (sample 427) showed limited cross-reactivity.

As an alternative method, genomic DNA in frozen tissues of lung cancer samples WAs extracted and BCR sequencing WAs performed using its facilitated sequencing and analysis method (IMMUNOSEQ Assay) (Adaptive biotechnology, Seattle, WA). This method identified 3548 heavy and 5913 light chain heavy and light chain sequences for the BCR in sample 463, and 5901 heavy and 10679 light chain sequences in sample 427. In each case, the two most abundant heavy and light chain clones were selected (FIG. 4A) and aligned to the IMGT BCR reference sequence using the V-QUEST tool. The alignment to the highest score (S) was recorded. For each rearranged sequence, the information on VDJ gene usage indicated by BCR sequencing results was verified by IMGT alignment results. The rearranged sequences were then extended to the full-length immunoglobulin sequences using the germline sequences of the validated V and J genes.

The heavy and light chains from the same sample were paired homogenously and expressed in 293T cells by transient transfection as shown in figure 4B, all eight of these combinations resulted in significant levels of antibody production, antibodies H5L and H6L 13 were selected and tested against 2L USC and 3L UAD samples, including the samples from which the antibody sequences were assembled, as shown in figure 4C, H6L showed strong binding to the cancer tissue from which the antibodies were obtained, it was interesting that H5L 12 showed not only binding to the L UAD tissue sample obtained from it, but also cross-reactivity to another L UAD tested, but no cross-reactivity to the 3 tested L USC samples.

The data presented herein demonstrate that it is possible to identify large numbers of immunoglobulin sequences based on de novo assembly of transcriptomes and targeted sequencing of genomic DNA isolated from clinical samples, and selected examples show exclusive specificity and broad cross-reactivity to cancer tissues. Several types of organization are worth considering.

First, because antibodies are assembled based on somatic and germline sequences, and because pairing of heavy and light chains is based on random combinations, the identified antibodies may not represent an endogenous antibody response to cancer. Thus, antibody discovery using the methods described herein may be better than describing endogenous antibody responses. Recent studies have shown that single cell based sequences can describe antibody responses (6).

In addition, the present invention provides methods for identifying antigens that are recognized by antibodies, such as H2L 7, that are capable of being used in the treatment of cancer, and that allow for the identification of antigens that are recognized by antibodies.

Third, it should be noted that although the depth and length of the RNAseq data in the TCGA database does not allow the assembly of full-length Ig sequences, the length and depth can be increased to obtain such sequences, as can some recent databases. It should be noted that the sequences reconstructed from CDR3 appear to produce broadly reactive antibodies, while the sequences reconstructed from targeted sequencing data include some somatic mutations and are therefore highly specific. These data motivate the possibility that somatic mutations that increase antibody affinity may also reduce cross-reactivity. If this hypothesis can be confirmed by further studies, antibody sequences reconstructed only from the CDR3 may be of intrinsic value for cancer diagnosis.

Taken together, the ability to generate antibodies with broad cross-reactivity or high specificity to cancer samples suggests that the platform described herein may allow the identification of fully human antibodies for the treatment and/or diagnosis of human cancer. For diagnosis, these antibodies can be further modified by mutagenesis to further eliminate their cross-reactivity with normal tissues. For therapeutic development, antibodies can be further modified to enhance effector function by Fc optimization (7), antibody-drug binding (8), bispecific antibodies (9), and chimeric antigen receptors (10).

Example 5 Process

Examples 1 to 4 utilize, but are not limited to, the following methods.

Accessing and analyzing TCGA data sets

BAM-formatted primary solid tumor RNA sequencing Data for 6 cancer types (glioblastoma multiforme, N: 154; low grade glioma, N: 512; lung adenocarcinoma, N: 525; lung squamous carcinoma, N: 474; pancreatic adenocarcinoma, N: 177; skin melanoma, N: 103) were obtained from the NCI Genomic Data Commons portal (https:// portal.gdc.cancer.gov /). First, all files are extracted into the fastq file using samtools (11) fastq function with-O settings. The reads were aligned to the reference V, D, J and C genes of the human B cell receptor by using the MIXCR (12) alignment function. Two rounds of contig assembly were then performed using the MIXCR rescue function in RNAseq mode. BCR CDR3 clones were assembled by using the MIXCR function based on aligned contigs. Further fragment usage and correlation analyses were performed on the BCR track using VDJtools (13) software and R software (https:// www.r-project. org /).

Calculation of IgH clonality

The IgH clonality is defined as the 1-Pielou homogeneity and effectively rearranged according to the following formula:

where Ci is the clone score of rearrangement i and N is the total number of rearrangements. Calculation of IgH clonality was achieved in the R software (https:// www.r-project. org /) using the vegetarian food drawer software package (14).

Genomic DNA extraction and BCR library sequencing

2 cases L UAD cancer samples (samples 463 and 429) and 3 cases L USC cancer samples (sample 427, sample 840, sample 350) were selected for DNA extraction using TRIZO L reagent (15596026, lot 86105, ThermoFisher Scientific) according to the manufacturer's instructions.

Recombinant antibody production

Each heavy chain was co-transfected with each light chain at a 1:1 ratio into HEK293T (CR L-11268, ATCC) using PO L YFECT Transfection Reagent (Qiagen, 1015586, batch 154044326) or L ipofectamine 3000(Invitrogen, L3000-015, batch 1811506) HEK293T cells were grown in DMEM (L ife Technologies, 11965-15142) supplemented with 10% fetal bovine serum (GE Healthcare L ife Sciences, SH30910.03) and 10ug/ml penicillin/streptomycin (GEHealthcare L ife Sciences, 0-122) three days after Transfection, supernatants were collected and the concentration of recombinant antibody expressed by the sandwich E L ISA was assessed using known standards.

Tissue, tissue microarray and immunogenic fluorescence assay

Frozen lung tumor samples were purchased from proteogenx (Inglewood, CA) frozen Tissue was embedded in a frozen mould (Tissue-Tek, 4728) together with o.c. t compound (Tissue-Tek, 4583) and cut into 5 micron sections frozen multi-Tissue microarrays (FMC402a) and lung Tissue microarrays (F L C320) were purchased from US Biomax, Inc (Rockville, MD) frozen Tissue sections and TMA were thawed at room temperature for 10-20 minutes and washed in PBS 3 times frozen sections were fixed in 4% formaldehyde at room temperature for 10 minutes, then sections were washed in PBS 3 times for 5 minutes each, then sections were incubated in blocking solution (5% normal goat serum in PBS) for 1 hour — sections were incubated with each recombinant antibody in complete DMEM at room temperature or 4 ℃ for 2 hours or 4 ℃ for 3 times, as above in PBS 3 times, and incubated with moist Tissue in blocking solution (thermo PBS) with Pro buffered saline, with anti-fisher P3 times, with Pro buffered saline, and with blocking buffer (3) in Pro buffered saline, e.g. 3 times buffered saline, with fluorescence buffered saline, e.g. 3.

Biometrics

The specific tests used to analyze each set of experiments are shown in the legend. Data were analyzed using a fisher exact test tabulation. Statistical calculations were performed using GraphPad Prism Software (GraphPad Software, San Diego, California).

Reference to the literature

1.Adams,G.P.&Weiner,L.M.Monoclonal antibody therapy ofcancer.Nat.Biotechnol.23,1147–1157(2005).

2.Weiner,L.,Murray,J.&Shuptrine,C.Antibody-based immunotherapy ofcancer:New insights,new targets.Cell 148,1081–1084(2012).

3.Li,B.et al.Landscape of tumor-infiltrating T cell repertoire ofhuman cancers.Nat.Genet.48,725-732(2016).

4.Linnemann,C.et al.Technical Reports High-throughput identificationof antigen-specific TCRs by TCR gene capture.Nat.Med.19,1534-1541(2013).

5.Dekosky,B.J.et al.High-throughput sequencing of the paired humanimmunoglobulin heavy and light chain repertoire.Nat.Biotechnol.31,166-199(2013).

6.Georgiou,G.et al.The promise and challenge of high-throughputsequencing of the antibody repertoire.Nat.Biotechnol.32,158-168(2014).

7.

P.,Jean-Mairet,J.,Moudry,R.,Amstutz,H.&Bailey,J.E.Engineeredglycoforms of an antineuroblastoma IgG1 with optimized antibody-dependentcellular cytotoxic activity.Nat.Biotechnol.17,176–80(1999).

8.Hurwitz,E.,Arnon,R.,Sahar,E.&Danon,Y.a Conjugate of Adriamycin andMonoclonal Antibodies To

Antigen Inhibits Human Neuroblastoma Cells inVitro.Ann.N.Y.Acad.Sci.417,125–136(1983).

9.Merchant,A.M.et al.An efficient route to human bispecificIgG.Nat.Biotechnol.16,677-681(1998).

10.Eshhar,Z.,Waks,T.,Gross,G.&Schindler,D.G.Specific activation andtargeting of cytotoxic lymphocytes through chimeric single chains consistingof antibody-binding domains and the gamma or zeta subunits of theimmunoglobulin and T-cell receptors.Proc.Natl.Acad.Sci.90,720-724(1993).

11.Li,H.et al.The Sequence Alignment/Map format andSAMtools.Bioinformatics 25,2078–2079(2009).

12.Bolotin,D.A.et al.MiXCR:software for comprehensive adaptiveimmunity profiling.Nat.Methods 12,380–381(2015).

13.Shugay,M.et al.VDJtools:Unifying Post-analysis of T Cell ReceptorRepertoires.PLoS Comput.Biol.11,1-16(2015).

Reuben, A.et al.TCR recombinant Intratum or specificity in L transformed L un induced adipogens an Association with Predicted Neoantigenic specificity and Postsurgical Recurrence cancer Discov.10, 1088-1097 (2017). Table 1-first 100 heavy chain CDRs 3 from patient 1

TABLE 2 first 100 heavy chain CDRs 3 from patient 1

TABLE 3 first 100 heavy chain CDRs 3 from patient 2

TABLE 4 first 100 heavy chain CDRs 3 from patient 2

TABLE 5 first 100 heavy chain CDRs 3 from patient 3

TABLE 6 first 100 heavy chain CDRs 3 from patient 3

TABLE 7-sample 350 (heavy chain)

TABLE 8-sample 350 (light chain)

TABLE 9-sample 427 (heavy chain)

TABLE 10 sample 427 (light chain)

TABLE 11 sample 429 (heavy chain)

TABLE 12 sample 429 (light chain)

TABLE 13-sample 463 (heavy chain)

TABLE 14-sample 463 (light chain)

TABLE 15-sample 840 (heavy chain)

TABLE 16-sample 840 (light chain)

TABLE 17 antibody sequences

Claims

1. A method of identifying one or more nucleic acid sequences encoding an antibody or antibody fragment from a transcriptome of a subject, the method comprising:

4. The method of claim 2, further comprising:

5. The method of any one of claims 1 to 4, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

7. The method of any one of claims 1 to 6, wherein the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating the clonality score of the antibody is determined at least in part by the formula:

8. The method of any one of claims 1 to 7, wherein said screening the nucleic acid sequences encoding one or more antibody fragments in the transcriptome of a series of control subjects comprises performing the FASTQ, MIXCR and vdjsols functions on the transcriptome data.

9. The method of any one of claims 1 to 8, wherein said screening the transcriptome of said subject for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences.

11. The method of any one of claims 1 to 10, wherein the sample is a tissue sample from a subject having or suspected of having a hyperproliferative disease.

12. The method of any one of claims 1 to 10, wherein the sample comprises one or more hyperproliferative cells selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

13. A computer-implemented method for detecting the presence of a nucleic acid sequence encoding an antibody or antibody fragment in a transcriptome of a sample, the method comprising: in a system comprising at least one processor and a memory,

b. calculating, by the at least one processor, a clonality score corresponding to one or more of the nucleic acid sequences in each of the first and second sets of nucleic acid sequences;

16. The method of claim 14, further comprising:

and wherein said step of identifying from the subject's transcriptome a nucleic acid sequence encoding at least one CDR of a variable chain is based on the step of comparing said plurality of sets of abundantly expressed nucleic acid sequences from step (h).

19. The method of any one of claims 13 to 18, wherein the screening step comprises screening for nucleic acid sequences encoding antibody fragments of one or more B Cell Receptor (BCR) sequences, and the step of calculating the clonality score of the antibody is determined at least in part by the formula:

20. The method of any one of claims 16 to 19, wherein the screening the transcriptome of a series of control subjects for nucleic acid sequences encoding one or more antibody fragments comprises performing FASTQ, MIXCR and vdjsols functions on the transcriptome data.

23. The method of any one of claims 13 to 22, wherein the sample is a tissue sample from a subject having or suspected of having a hyperproliferative disease.

24. The method of any one of claims 13 to 23, wherein the sample comprises one or more hyperproliferative cells selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

27. A method of designing an antibody or antibody fragment capable of binding a cellular epitope from a sample, the method comprising:

(c) cloning one or more nucleic acids in a vector or synthesizing the antibody by solid state chemical synthesis; and

if the one or more nucleic acid sequences are cloned into a vector, then (d) the vector is transformed into a host cell, and (e) sufficient time is allowed for the recombinant production of the encoded antibody or antibody fragment by the host cell.

28. A composition comprising an antibody or antibody fragment comprising at least one CDR sequence obtained by performing the method of any one of claims 1 to 25.

29. An antibody library comprising at least one amino acid sequence obtained by performing the method of any one of claims 1 to 27.

30. A non-transitory computer program product encoded on a computer-readable storage medium, the non-transitory computer program product comprising instructions for:

31. The non-transitory computer program product of claim 30, further comprising instructions for: (d) after performing the step of classifying, identifying from the first set of nucleic acid sequences a nucleic acid sequence encoding at least one Complementarity Determining Region (CDR) of the variable strand.

32. The non-transitory computer program product of claim 31, further comprising instructions for repeating the steps of identifying a nucleic acid sequence encoding at least one CDR of a variable chain and editing a plurality of CDR sequences.

33. The non-transitory computer program product of claim 31, further comprising instructions for:

34. The non-transitory computer program product of claim 31, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of antibody fragments selected from the group consisting of: an Fc portion of an antibody, a single chain variable fragment of an antibody (ScFv), an Fv portion of an antibody, an Fab fragment of an antibody, an F (ab')2 fragment of an antibody, an Fd fragment of an antibody, an IgG-like fragment of an antibody, a variable chain of an antibody, and a constant region of an antibody.

35. The non-transitory computer program product of claims 30-34, wherein the first set of abundantly expressed nucleic acid sequences from the subject encodes one or a combination of variable heavy and/or variable light chain portions of the antibody.

36. The non-transitory computer program product of any one of claims 30 to 35, wherein the screening step comprises screening nucleic acid sequences of antibody fragments encoding one or more B Cell Receptor (BCR) sequences, and the step of calculating a clonality score for an antibody is determined at least in part by the formula:

37. The non-transitory computer program product of claims 30-36, wherein the screening nucleic acid sequences encoding one or more antibody fragments in the transcriptome of a series of control subjects comprises performing FASTQ, MIXCR, and vdjools functions on the transcriptome data.

38. The non-transitory computer program product of claims 30-37, wherein the screening the transcriptome of the subject for nucleic acid sequences encoding one or more antibody fragments comprises performing an alignment of cloned sequences.

39. The non-transitory computer program product of claim 38, wherein the alignment of cloned sequences comprises performing an immunizing SEQ function.

40. The non-transitory computer program product of any one of claims 30 to 39, wherein the sample is a tissue sample from a subject having a hyperproliferative cellular disease.

41. The non-transitory computer program product of claims 30-40, wherein the sample is a cancer tissue selected from the following cancers: glioblastoma multiforme, low-grade gliomas, lung adenocarcinomas, lung squamous carcinomas, pancreatic adenocarcinomas and skin cancers.

42. An antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising:

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2206; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2208; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence shown in SEQ ID NO: 2210; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO 2216; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2218; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO 2220;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO. 2226; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO. 2228; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 2230; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2236; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2238; and a light chain CDR3, said light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2240;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2246; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2248; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO 2250; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO 2256; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO 2258; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2260;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2266; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2268; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2270; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2276; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2278; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2280;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO. 2286; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO 2288; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2290; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2296; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO 2298; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2300;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2306; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2308; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 2310; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2316; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2318; and a light chain CDR3, the light chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2320;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2326; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2328; a heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2330; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2336; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2338; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2340;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 2346; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 2348; a heavy chain CDR3, the heavy chain CDR3 includes an amino acid sequence shown in SEQ ID NO. 2350; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2356; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2358; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2360;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2366; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2368; a heavy chain CDR3, said heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 2370; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2376; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2378; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2380;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2386; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2388; a heavy chain CDR3, said heavy chain CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 2390; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2396; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2398; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2400;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2406; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2408; a heavy chain CDR3, the heavy chain CDR3 comprises an amino acid sequence shown as SEQ ID NO: 2410; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2416; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2418; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO. 2420;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2426; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2428; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 2430; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2436; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2438; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2440;

a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2446; a heavy chain CDR2, the heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2448; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2450; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2456; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2458; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2460;

a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2466; a heavy chain CDR2, the heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2468; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2470; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2476; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2478; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2480;

a heavy chain CDR1, the heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2486; a heavy chain CDR2, the heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2488; a heavy chain CDR3, the heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2490; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2496; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2498; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2500;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2506; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2508; a heavy chain CDR3, wherein the heavy chain CDR3 comprises an amino acid sequence shown as SEQ ID NO: 2510; a light chain CDR1, said light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2516; a light chain CDR2, said light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2518; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2520;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence as shown in SEQ ID NO: 2526; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence as shown in SEQ ID NO: 2528; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2530; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2536; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2538; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2540;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2546; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2548; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2550; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2556; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2558; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2560;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2566; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2568; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2570; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2576; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2578; and a light chain CDR3, the light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2580;

a heavy chain CDR1, said heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2586; a heavy chain CDR2, said heavy chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2588; a heavy chain CDR3, said heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO. 2590; a light chain CDR1, the light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 2596; a light chain CDR2, the light chain CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 2598; and a light chain CDR3, said light chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 2600;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2606; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2608; a heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2610; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2616; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2618; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2620;

a heavy chain CDR1, said heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2626; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2628; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence shown in SEQ ID NO: 2630; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2636; a light chain CDR2, the light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2638; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2640;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2646; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2648; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence shown in SEQ ID NO: 2650; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2656; a light chain CDR2, the light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2658; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2660;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2666; a heavy chain CDR2, said heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2668; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence shown in SEQ ID NO: 2670; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2676; a light chain CDR2, the light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2678; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2680;

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2686; heavy chain CDR2, said heavy chain CDR2 includes the amino acid sequence shown in SEQ ID NO: 2688; a heavy chain CDR3, said heavy chain CDR3 comprising the amino acid sequence shown in SEQ ID NO: 2690; a light chain CDR1, the light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2696; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2698; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 2700; and

heavy chain CDR1, said heavy chain CDR1 includes the amino acid sequence shown in SEQ ID NO: 2706; heavy chain CDR2, said heavy chain CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 2708; a heavy chain CDR3, said heavy chain CDR3 includes the amino acid sequence shown in SEQ ID NO: 2710; a light chain CDR1, said light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 2716; a light chain CDR2, said light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2718; and a light chain CDR3, the light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO 2720.

43. An antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising:

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID No. 2204 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2204, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID No. 2214 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 2214;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2224 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2224, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2234 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2234;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2244 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2244, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2254 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2254;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2264 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2264, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2274 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2274;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2284 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2284, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2294 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2294;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2304 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2304, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2314 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2314;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2324 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2324, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2334 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2334;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2344 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2344, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2354 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2354;

a heavy chain variable domain comprising the amino acid sequence set forth as SEQ ID NO:2364 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2364, and/or a light chain variable domain comprising the amino acid sequence set forth as SEQ ID NO:2374 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2374;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2384 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2384, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2394 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2394;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2404 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2404, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2414 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2414;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2424 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2424, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2434 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2434;

a heavy chain variable domain comprising the amino acid sequence set forth as SEQ ID NO:2444 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2444, and/or a light chain variable domain comprising the amino acid sequence set forth as SEQ ID NO:2454 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2454;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2464 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2464, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2474 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2474;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2484 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2484, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2494 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2494;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2504 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2504, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2514 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2514;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2524 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2524, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2534 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2534;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2544 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2544, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2554 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2554;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2564 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2564, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2574 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2574;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2584 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO:2584, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2594 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 2594;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2604 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2604, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2614 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2614;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2624 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2624, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2634 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2634;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2644 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2644, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2654 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2654;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2664 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2664, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2674 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2674;

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2684 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2684, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2694 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2694; and

a heavy chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2704 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2704, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2714 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2714.

44. An antibody or antigen-binding fragment thereof that binds to tumor tissue, comprising:

a heavy chain comprising the amino acid sequence shown as SEQ ID No. 2202 or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 2202, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID No. 2212 or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 2212;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2222 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2222, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2232 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2232;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2242 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2242, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2252 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2252;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2262 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2262, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2272 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2272;

a heavy chain comprising the amino acid sequence set forth as SEQ ID NO. 2282 or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO. 2282, and/or a light chain variable domain comprising the amino acid sequence set forth as SEQ ID NO. 2292 or a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO. 2292;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2302 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2302, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2312 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2312;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2322 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2322, and/or a light chain, said light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2332 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2332;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2342 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2342, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2352 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2352;

a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2362 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2362, and/or a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:2372 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2372;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2382 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2382, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2392 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2392;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2402 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2402, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2412 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2412;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2422 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2422, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2432 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2432;

a heavy chain comprising the amino acid sequence set forth as SEQ ID NO:2442 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2442, and/or a light chain variable domain comprising the amino acid sequence set forth as SEQ ID NO:2452 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2452;

a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2462 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2462, and/or a light chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:2472 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2472;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2482 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2482, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2492 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2492;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2502 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2502, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2512 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2512;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2522 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2522, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO:2532 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2532;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2542 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2542, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2552 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2552;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2562 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2562, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2572 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2572;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2582 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2582, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO. 2592 or a sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO. 2592;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2602 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2602, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2612 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2612;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2622 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2622, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2632 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2632;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2642 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2642, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2652 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2652;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2662 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2662, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2672 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2672;

a heavy chain comprising the amino acid sequence shown as SEQ ID NO:2682 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:2682, and/or a light chain variable domain comprising the amino acid sequence shown as SEQ ID NO:2692 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 2692; and

a heavy chain comprising the amino acid sequence shown as SEQ ID NO. 2702 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2702, and/or a light chain comprising the amino acid sequence shown as SEQ ID NO. 2712 or a sequence having at least 90%, 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO. 2712.

45. The antibody or antigen-binding fragment thereof that binds to tumor tissue according to claim 44, wherein

The heavy chain comprises an amino acid sequence shown as SEQ ID NO:2202, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2212;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO. 2222, and the light chain comprises an amino acid sequence shown as SEQ ID NO. 2232;

the heavy chain comprises an amino acid sequence as shown in SEQ ID NO 2242 and the light chain comprises an amino acid sequence as shown in SEQ ID NO 2252;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2262, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2272;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO. 2282, and the light chain comprises the amino acid sequence shown as SEQ ID NO. 2292;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO. 2302 and the light chain comprises the amino acid sequence shown as SEQ ID NO. 2312;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2322, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2332;

the heavy chain comprises an amino acid sequence as set forth in SEQ ID NO:2342 and the light chain comprises an amino acid sequence as set forth in SEQ ID NO: 2352;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2362, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2372;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2382, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2392;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2402, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2412;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2422, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2432;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2442, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2452;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2462 and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2472;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2482 and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2492;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2502, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2512;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO:2522, and the light chain comprises an amino acid sequence shown as SEQ ID NO: 2532;

the heavy chain comprises an amino acid sequence shown as SEQ ID NO. 2542, and the light chain comprises an amino acid sequence shown as SEQ ID NO. 2552;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO. 2562, and the light chain comprises the amino acid sequence shown as SEQ ID NO. 2572;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO. 2582, and the light chain comprises the amino acid sequence shown as SEQ ID NO. 2592;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2602, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2612;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2622 and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2632;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2642, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2652;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2662, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2672;

the heavy chain comprises the amino acid sequence shown as SEQ ID NO:2682, and the light chain comprises the amino acid sequence shown as SEQ ID NO: 2692; or

The heavy chain comprises the amino acid sequence shown as SEQ ID NO 2702 and the light chain comprises the amino acid sequence shown as SEQ ID NO 2712.

46. The antibody or antigen-binding fragment thereof of any one of claims 42 to 45, wherein the fragment is selected from the group consisting of Fab, Fab ', F (ab')2, Fv, domain antibody, and single chain antibody.

47. The antibody or antigen-binding fragment thereof of any one of claims 42-46, wherein the antibody or antigen-binding fragment thereof is classified as an isotype selected from the group consisting of IgG, IgM, IgD, IgA, and IgE.

48. An antibody or antigen-binding fragment thereof that binds to tumor tissue, which competes with the antibody or antigen-binding portion thereof of any one of the preceding claims.

49. The antibody or antigen-binding fragment thereof of any one of claims 42 to 48, wherein the antibody or antigen-binding portion thereof is humanized.

50. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 42 to 49, and a pharmaceutically acceptable carrier.

51. The pharmaceutical composition of claim 50, which is lyophilized.

52. An isolated nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 42 to 48.

53. A vector comprising the isolated nucleic acid of claim 52.

54. A host cell comprising the vector of claim 53.

55. The host cell of claim 54, wherein the host cell is a prokaryotic cell or a eukaryotic cell.

56. The host cell of claim 55, wherein the eukaryotic cell is a protoplast cell, an animal cell, a plant cell, a fungal cell, a yeast cell, a mammalian cell, an avian cell, or an insect cell.

57. The host cell of claim 56, wherein the mammalian cell is a CHO cell or a COS cell.