RU2785291C2 - Methods and compositions for prediction of therapeutic efficiency of treatment of oncological diseases and prediction of oncological diseases - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to methods for determination of whether a patient with an oncological disease, having tumor positive by a tumor antigen, responds to treatment, using an antibody against a tumor antigen, and methods for determination of whether a patient with an oncological disease, having tumor positive by a tumor antigen, will have a progression-free survival period. In this case, the tumor antigen is CLDN18.2 protein. A method for the treatment of a patient with an oncological disease, having tumor positive by an antigen, is also presented.

EFFECT: invention allows for its effective use for prediction of the efficiency of treatment of malignant neoplasms and prediction of clinical outcomes for patients with oncological diseases.

16 cl, 14 dwg, 16 tbl, 3 ex

Description

Technical field

The invention generally relates to methods and compositions for predicting the therapeutic efficacy of cancer treatment and cancer prognosis. The invention discloses markers that are associated with favorable and unfavorable outcomes, respectively, in certain cancer treatments and are useful as cancer prognostic markers. Methods associated with these markers are disclosed for predicting the effectiveness of cancer treatment and predicting clinical outcomes for cancer patients.

Prior Art

Cancer of the stomach and esophagus (gastroesophageal, GE) is one of the malignant neoplasms with the highest unsatisfied medical demand. Stomach cancer is the second leading cause of death worldwide. The incidence of esophageal cancer has increased in recent decades, and the overall five-year survival rate for GI cancer is 20-25%, despite the aggressiveness of the established standard of care associated with significant side effects. The demand for medical care in patients suffering from this type of malignant neoplasms is high, and innovative drugs are required.

Tight junction molecule claudin 18, isotype 2 (CLDN18.2) is a cancer-associated splicing variant of claudin 18 [Niimi, T., et al., Mol Cell Biol, 2001. 21 (21): p. 7380-90; Tureci, O., et al., Gene, 2011. 481(2): p. 83-92]. CLDN18.2 is a 27.8 kDa transmembrane protein containing four transmembrane domains with two small extracellular loops (loop 1 spanned by hydrophobic region 1 and hydrophobic region 2, loop 2 spanned by hydrophobic regions 3 and 4). CLDN18.2 is a highly selective gastric-derived antigen exclusively expressed on short-lived differentiated gastric epithelial cells and not found in any other normal human tissue. The antigen is ectopically expressed at significant levels in a variety of human malignancies, including gastroesophageal cancer and pancreatic cancer [Sahin, U., et al., Clin Cancer Res, 2008. 14 (23): p. 7624-34]. The CLDN18.2 protein is also commonly found in lymph node metastases from gastric cancer and in distant metastases. CLDN18.2 appears to be involved in the proliferation of CLDN18.2-positive tumor cells, as downregulation of the target by siRNA technology results in inhibition of gastric cancer cell proliferation.

IMAB362 is a chimeric IgG1 monoclinal antibody directed against CLDN18.2. IMAB362 recognizes the first extracellular domain of CLDN18.2 with high affinity and specificity and does not bind to any other member of the Claudio family, including the closely related claudin 18 splice variant 1 (CLDN18.1).

In mice bearing human xenografts expressing CLDN18.2, after administration of IMAB362, an increase in survival and regression of tumors was observed. When intravenously administered to relevant animal species, toxicity in gastric tissue was not observed, since the target epitope is not available. However, the tumor target becomes available to IMAB362 during malignant transformation. IMAB362 combines four independent potent mechanisms of action: (i) antibody-dependent cellular cytotoxicity (ADCC), (ii) complementary-dependent cytotoxicity (CDC), (iii) induction of apoptosis caused by cross-linking of a target on the tumor surface, and (iv) direct inhibition of proliferation.

A previous phase I study evaluated IMAB362 as single dose monotherapy in patients with advanced gastroesophageal cancer. Five doses of IMAB362 (33, 100, 300, 600 and 1000 mg/m²) were used as monotherapy in this study. This study shows that a single dose of this antibody is safe and well tolerated at doses up to 1000 mg/m², as there were no significant differences in AE profile and other safety parameters between dose groups (AE = adverse event). The best results in terms of antitumor activity were obtained for the 300 mg/m² and 600 mg/m² groups. Two patients in the 300 mg/m2 group were disease controlled and because they had only off-target lesions, they were scored as non-CR, non-PD (CD=complete response, PD=progressive disease). The duration of non-CR, non-PD was about two months and six weeks, respectively. The levels of tumor markers of these three patients remained stable. One patient in the 600 mg/m² group experienced stable disease (SD). The duration of SD was about 2 months.

Based on the potent mechanisms of action for IMAB362-induced cell killing, the survival advantage of IMAB362-treated mice bearing CLDN18.2-positive tumors, the absence of any evidence of IMAB362-associated toxicity, and the promising results of the Phase I study, a Phase IIa study was initiated. This Phase IIa clinical study was conducted to determine the safety, tolerability, and antitumor activity of repeated doses of IMAB362 in patients with histologically proven metastatic, refractory, or recurrent gastric or lower esophageal advanced adenocarcinoma disease.

In this phase IIa study, the study drug was used in three cohorts that were recruited sequentially. The first cohort of three patients received repeated doses of IMAB362 at a lower dose level (300 mg/m2 body surface area). The antibody was administered as an intravenous infusion over 2 hours. Since there were no signs of toxicity associated with IMAB362 in the first cohort, the dose of IMAB362 in the second cohort (three patients) was increased to 600 mg/m2 body surface area. In the third cohort, 19 patients were assigned to the same dose (repetitive application of 600 mg/m2 body surface area). Patient samples from this cohort were analyzed for several concomitant analyses, eg ADCC, CDC, immunophenotyping, and genetic immune polymorphisms. All patients in all cohorts received repeat doses of IMAB362 every other week at visits 2, 5, 6, 7 and 8 (5 applications).

The divergence of antigen-positive tumors (overexpressing the target antigen to the same extent) in terms of responsiveness to intervention with therapeutic monoclonal antibodies such as IMAB362 indicates the presence of additional factors related to the outcome of therapy. This requires careful selection of patients who may benefit from antibody therapy.

Therefore, it is necessary to develop a test to assess the acceptability of patients for antibody therapy. The present invention satisfies this need by providing markers that are associated with favorable and unfavorable outcomes, respectively, in antibody therapy. In addition, the present invention demonstrates that these markers are useful as markers for predicting clinical outcomes for cancer patients.

The results presented herein can be used to select the appropriate treatment for a cancer patient and, in particular, to decide whether to administer antibody therapy to a cancer patient.

The essence of the invention

The present invention relates to methods for genotyping SNPs (single nucleotide polymorphisms), for example, for use in assessing the likelihood of an individual to respond to therapeutic treatment for cancer, in choosing a treatment or prophylaxis regimen (for example, in deciding whether to administer a particular therapeutic agent to an individual, having cancer, or who has an increased risk of developing cancer in the future), or when evaluating an individual's prognosis for disease severity and recovery.

The present invention is based on the fact that certain SNP genotypes are associated with cancer sensitivity/insensitivity to antibody treatment, such as treatment of CLDN18.2 positive cancer, in particular CL1318.2 positive gastroesophageal cancer with IMAB362. The present invention is also based on the fact that certain SNP genotypes are associated with clinical outcome for cancer patients and thus are useful in cancer prognosis.

In one aspect, the invention relates to a method for evaluating

(i) if a cancer patient with a tumor antigen positive tumor is a responder to treatment with an anti-tumor antigen antibody and/or

(ii) if a cancer patient, preferably a cancer patient, having a tumor antigen positive tumor will have progression-free survival,

причем указанный способ включает определение генотипа для одного или нескольких однонуклеотидных полиморфизмов, выбранных из группы, состоящей из FCGR2A rs1801274, MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260, ERCC1 rs11615 и FCGR3A rs396991 в образце, obtained from the patient.

In one embodiment, the presence of the heterozygous FCGR2A rs1801274 [CT] genotype is indicative of a reduced risk that the cancer patient will not respond to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous FCGR2A rs1801274 [TT] genotype and/or the homozygous FCGR2A rs1801274 [CC] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient the disease does not have a progression-free survival period.

In one embodiment, the presence of the homozygous MUC1 rs4072037 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous IL-10 rs1800896 [GG] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a heterozygous DNMT3A rs1550117 [GA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the heterozygous SMAD4 rs12456284 [GA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous EGF rs4444903 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a homozygous CDH1 rs16260 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a homozygous ERCC1 rs11615 [TT] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a heterozygous FCGR3A rs396991 [TG] genotype and/or a homozygous FCGR3A rs396991 [TT] genotype is indicative of a reduced risk that a cancer patient is not a responder to antibody treatment and/or a reduced risk that a cancer patient the disease does not have a progression-free survival period.

In one embodiment, the presence of the homozygous FCGR3A rs396991 [GG] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the tumor antigen is a CLDN18.2 protein.

In one aspect, the invention relates to a method for evaluating

(i) if a cancer patient with a CLDN18.2 positive tumor is a responder to treatment with an antibody against the CLDN18.2 protein and/or

(ii) if a cancer patient, preferably a cancer patient, having a CLDN18.2 positive tumor will experience progression-free survival,

причем указанный способ включает определение генотипа для одного или нескольких однонуклеотидных полиморфизмов, выбранных из группы, состоящей из FCGR2A rs1801274, MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260, ERCC1 rs11615 и FCGR3A rs396991 в образце, obtained from the patient.

In one embodiment, the presence of the heterozygous FCGR2A rs1801274 [CT] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous FCGR2A rs1801274 [TT] genotype and/or the homozygous FCGR2A rs1801274 [CC] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient there will be no progression-free survival.

In one embodiment, the presence of the homozygous MUC1 rs4072037 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous IL-10 rs1800896 [GG] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a heterozygous DNMT3A rs1550117 [GA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the heterozygous SMAD4 rs12456284 [GA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of the homozygous EGF rs4444903 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a homozygous CDH1 rs16260 [AA] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a homozygous ERCC1 rs11615 [TT] genotype is indicative of a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period.

In one embodiment, the presence of a heterozygous FCGR3A rs396991 [TG] genotype and/or a homozygous FCGR3A rs396991 [TT] genotype is indicative of a reduced risk that a cancer patient is not a responder to antibody treatment and/or a reduced risk that a cancer patient the disease does not have a progression-free survival period.

In one embodiment, the presence of a homozygous FCGR3A rs396991 [GG] genotype indicates an increased risk that the cancer patient will not respond to antibody treatment and/or an increased risk that the cancer patient will not have a progression-free survival period.

In one embodiment of all aspects of the invention, the antibody acts by recruiting the patient's immune system to destroy tumor cells. In one embodiment, the antibody acts through antibody dependent cell mediated cytotoxicity (ADCC) and/or complement dependent cytotoxicity (CDC). In one embodiment, the antibody is a monoclonal antibody. In one embodiment of all aspects of the invention, the antibody comprises a heavy chain containing the amino acid sequence represented by SEQ ID NO: 17 or 51, or a fragment thereof, and a light chain containing the amino acid sequence represented by SEQ ID NO: 24, or a fragment thereof.

In one embodiment of all aspects of the invention, resistance to antibody treatment includes a relative reduction in one or more of survival, progression-free survival, disease-free survival, long-term disease-free survival, and stable disease.

In one aspect, the invention relates to a method for treating a patient with cancer, said method comprising

a. evaluating whether a cancer patient is a responder to treatment with an antibody according to the method of the invention, and

b. (i) treating a cancer patient with an antibody if the patient has a reduced risk of not being a responder to antibody treatment, or (ii) not treating a cancer patient with an antibody and/or treating a cancer patient with a treatment regimen that includes treatment that differs from antibody treatment if the patient is at increased risk of not responding to antibody treatment.

In one embodiment, the treatment regimen includes treatment independent of the patient's immune system. In one embodiment, the treatment regimen does not include treatment with an antibody that acts by recruiting the patient's immune system to kill tumor cells. In one embodiment, the treatment regimen includes surgery, chemotherapy and/or radiation. In one embodiment, the treatment regimen includes treatment with a small molecule inhibitor of a tumor antigen and/or an antibody-drug conjugate, wherein the antibody is directed against the tumor antigen. In one embodiment, the antibody-drug conjugate is an antibody linked to a radioactive, chemotherapeutic, or toxic moiety. In one embodiment, the antibody-drug conjugate is an antibody linked to a cytostatic or cytotoxic compound.

In one aspect, the invention relates to a method for assessing clinical outcome for a patient with cancer, which method includes determining the genotype for one or more single nucleotide polymorphisms selected from the group consisting of FCGR2A rs1801274, MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260, ERCC1 rs11615 and FCGR3A rs396991 in a patient sample.

In one embodiment, the presence of a heterozygous FCGR2A rs1801274 [CT] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a homozygous FCGR2A rs1801274 [TT] genotype and/or a homozygous FCGR2A rs1801274 [CC] genotype indicates an increased risk of a poor clinical outcome.

In one embodiment, the presence of a homozygous MUC1 rs4072037 [AA] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk of adverse clinical outcome.

In one embodiment, the presence of a homozygous IL-10 rs1800896 [GG] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a heterozygous DNMT3A rs1550117 [GA] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of the heterozygous SMAD4 rs12456284 [GA] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of the homozygous EGF rs4444903 [AA] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a homozygous CDH1 rs16260 [AA] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a homozygous ERCC1 rs11615 [TT] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of a heterozygous FCGR3A rs396991 [TG] genotype and/or a heterozygous FCGR3A rs396991 [TT] genotype is indicative of a reduced risk of adverse clinical outcome.

In one embodiment, the presence of the homozygous FCGR3A rs396991 [GG] genotype is indicative of an increased risk of adverse clinical outcome.

In one embodiment, assessing a clinical outcome for a cancer patient includes predicting the likelihood of one or more of survival, progression-free survival, disease-free survival, long-term disease-free survival, and stable disease. In one embodiment, the adverse clinical outcome includes a relative decrease in one or more of survival, progression-free survival, disease-free survival, long-term disease-free survival, and stable disease.

In one embodiment, the patient has a tumor antigen positive tumor and is being treated with an antibody against the tumor antigen.

In one embodiment of all aspects of the invention, the sample is a sample containing DNA. In one embodiment, DNA has been extracted from a sample of a patient's body. In one embodiment, DNA is extracted from blood.

In one embodiment of all aspects of the invention, the tumor is a solid tumor. In one embodiment, the tumor is a gastroesophageal tumor. In one embodiment, the tumor is a progressive adenocarcinoma of the stomach or lower esophagus. In one embodiment, the cancer is gastroesophageal cancer. In one embodiment, the cancer is a progressive adenocarcinoma of the stomach or lower esophagus.

In a further aspect, the present invention relates to a kit containing an agent for determining the genotype of one or more single nucleotide polymorphisms selected from the group consisting of FCGR2A rs1801274, MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF Rs4444903, rs11615 and FCGR3A rs396991 in a patient sample. In one embodiment, said kit is useful for carrying out the methods of all aspects of the present invention. In one embodiment, said set further comprises a storage medium. In one preferred embodiment, said storage medium is an electronic or non-electronic storage medium. In one embodiment, said storage medium contains instructions on how to perform the methods of all aspects of the invention.

Other objects, advantages and features of the present invention will become apparent from the following detailed description when considered in conjunction with the accompanying drawings.

Brief description of the drawings

Fig. 1: Single nucleotide polymorphisms with a statistically significant genotype frequency shift between patient and control population (χ ² test, p<0.05).

SNP genotypes are indicated in the bar sections. Pat. Patient population, Co. control population.

Fig. 2: Relative frequency of homozygous risk genotypes per patient relative to the number of SNP risk factors studied per patient. Patients are sorted by increasing frequency of accumulated homozygous risk factors.

Fig. 3: Progression-free survival of PP patients differentiated by the rs1801274 (FCGR2A) genotype (Kaplan-Meier curve)

Fig. 4: Progression-free survival of FAS patients differentiated by the rs1801274 (FCGR2A) genotype (Kaplan-Meier curve)

Fig. 5: Progression-free survival of PP patients differentiated by the rs1800896 (IL-10) genotype (Kaplan-Meier curve)

Fig. 6: Progression-free survival of FAS patients differentiated by the rs1800896 (IL-10) genotype (Kaplan-Meier curve)

Fig. 7: Progression-free survival of FAS patients differentiated by the rs1550117 (DNMT3A) genotype (Kaplan-Meier curve)

Fig. 8: Progression-free survival of PP patients differentiated by the rs12456284 (SMAD4) genotype (Kaplan-Meier curve)

Fig. 9: Progression-free survival of PP patients differentiated by the rs4072037 (MUC1) genotype (Kaplan-Meier curve)

Fig. 10: Progression-free survival of FAS patients differentiated by the rs4072037 (MUC1) genotype (Kaplan-Meier curve)

Fig. 11: Progression-free survival of FAS patients differentiated by the rs4444903 (EGF) genotype (Kaplan-Meier curve)

Fig. 12: Progression-free survival of FAS patients differentiated by the rs16260 (CDH1) genotype (Kaplan-Meier curve)

Fig. 13: Progression-free survival of PP patients differentiated by the rs11615 (ERCC1) genotype (Kaplan-Meier curve)

Fig. 14: Progression-free survival of PP patients differentiated by the rs396991 (FCGR3A) genotype (Kaplan-Meier curve).

Detailed description of the invention

Although the present invention is described in detail below, it should be understood that this invention is not limited to the specific methodologies, protocols, and reagents described herein, as they may vary. In addition, it should be understood that the terminology used herein is for the purpose of describing specific embodiments only, and is not intended to be limiting, as the scope of the present invention will only be limited by the appended claims. Unless otherwise indicated, all technical and scientific terms used in this document have the same meanings as generally understood by a person skilled in the art.

The elements of the present invention will be described below. These elements are listed in specific embodiments, however, it should be understood that they can be combined in any way and in any number to create additional embodiments. The various described examples and preferred embodiments should not be construed as limiting the present invention to the expressly described embodiments. This description is to be understood as supporting and covering embodiments that combine the expressly described embodiments with any number of disclosed and/or preferred elements. In addition, any permutations and combinations of all described elements in this application should be considered as disclosed by the description of this application, unless the context indicates otherwise.

Preferably, the terms used in this document are defined in accordance with the description "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)", HGW Leuenberger, B. Nagel and H. Kölbl, Eds., Helvetica Chimica Acta, CH-4010 Basel, Switzerland, (1995).

The practice of the present invention will use, unless otherwise noted, the conventional methods of chemistry, biochemistry, cell biology, immunology, and recombinant DNA as explained in the literature in the art (see, for example, Molecular Cloning: A Laboratory Manual, ^2nd Edition, J. Sambrook et al.eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989).

Throughout this description and the following claims, unless the context otherwise requires, the word "comprise" and variations such as "comprises" and "comprising" will be understood to include the claimed element, whole or step or group of members, whole or steps , but not the exclusion of any member, whole or stage or group of members, integers or stages, although in some embodiments such other element, whole or stage or group of members, integers or stages, i.e. the object of the invention is to include the specified member, whole or stage or group of members, integers or stages. The terms "a" and "an" and "the" and similar references used in the context of the description of the invention (especially in the context of the claims) should be construed as covering both the singular and the plural, unless otherwise specified in this document or not. clearly inconsistent with the context. The presentation of ranges of values in this document is simply intended to be used as a shorthand way of referring individually to each individual value within a range. Unless otherwise noted, each individual value is included in the description as if it were individually described in this document. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or otherwise clearly contrary to the context. The use of any and all examples or exemplary language (eg, "such as") provided herein is merely intended to better illustrate the invention and does not constitute a limitation on the scope of the invention as otherwise claimed. No wording in the specification should be construed as indicating any unclaimed element as being essential in the practice of the invention.

Several documents are cited in the text of this description. Each of the documents cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether above or below, is incorporated herein by reference in its entirety. No part of this document should be taken as an admission that the invention cannot claim an earlier date for the present description by virtue of prior invention.

The present inventors provide tests for evaluating the acceptability of patients for certain cancer therapies, in particular antibody therapy, and for inferring the prognosis of a patient with cancer. The results obtained using these tests allow the physician to select the appropriate treatment for the cancer patient and in particular to decide whether to administer antibody therapy to a particular cancer patient.

The term "single nucleotide polymorphism" or "SNP" refers to a DNA sequence variation commonly found in a population in which one nucleotide in the genome (or other common sequence) differs between members of a species or paired chromosomes. SNPs can occur in the coding sequences of genes, non-coding regions of genes, or in intergenic regions (regions between genes). SNPs within a coding sequence may, but need not, change the amino acid sequence of the protein produced due to the degeneracy of the genetic code. Thus, SNPs in the coding region are of two types: synonymous and non-synonymous SNPs. Synonymous SNPs do not affect the protein sequence, while nonsynonymous SNPs change the amino acid sequence of the protein. Nonsynonymous SNPs are of two types: missense and meaningless (nonsense). SNPs that are not in protein-coding regions can still affect gene splicing, transcription factor binding, messenger RNA degradation, or non-coding RNA sequencing. Gene expression that is affected by this type of SNP is referred to as eSNP (expression SNP) and such SNPs can be upstream or downstream of a gene in sequence.

Various methods known in the art can be used to determine the genotype of an SNP. Analytical methods for detecting novel SNPs and detecting known SNPs include, for example, DNA sequencing, capillary electrophoresis, mass spectrometry, single strand conformational polymorphism (SSCP), electrochemical analysis, denaturing HPLC and gel electrophoresis, restriction fragment length polymorphism, and hybridization analysis.

The process of determining which nucleotide is present at a particular SNP position described herein for one or both alleles may be referred to by phrases such as "SNP genotyping" or "SNP genotyping". Thus, these phrases may refer to the detection of a single allele (nucleotide) at a SNP position, or may encompass the detection of both alleles (nucleotides) at a SNP position (eg, to determine whether a SNP position is homozygous or heterozygous). In addition, these phrases may also refer to the discovery of the amino acid residue encoded by the SNP (such as alternative amino acid residues that are encoded by different codons created by alternative nucleotides at the position of the SNP).

A reagent that specifically detects a particular position of a target SNP as disclosed herein, and which preferably is specific for a particular nucleotide (allele) of the position of the target SNP (i.e., the reagent can preferably distinguish between different alternative nucleotides at the position of the target SNP, thereby allowing nucleotide identity to be determined, present at a given SNP position) can be used to detect the SNP. Typically, such a detection reagent hybridizes to a nucleic acid molecule containing the target SNP by complementary base pairing in a specific manner and distinguishes the target variant sequence from other nucleic acid sequences, such as a known form in the art in the test sample. An example of a detection reagent is a non-natural nucleotide primer or probe that hybridizes to a target nucleic acid containing the SNP disclosed herein. In a preferred embodiment, such a primer or probe can distinguish nucleic acids having a particular nucleotide (allele) at the position of the target SNP from other nucleic acids that have a different nucleotide at the same position of the target SNP. In addition, the detection reagent may hybridize to a specific region 5' and/or 3' relative to the position of the SNP. One of skill in the art will appreciate that such detection reagents, eg, primers and probes, are directly useful as reagents for genotyping one or more of the SNPs described herein and may be included in any kit format.

For SNP analysis, it may be useful to use oligonucleotides specific for alternative SNP alleles. Such oligonucleotides that show single nucleotide variations in target sequences may be referred to by terms such as "allele-specific oligonucleotides", "allele-specific probes", or "allele-specific primers".

The SNP detection reagent may be labeled with a reporter, such as a fluorographic reporter dye, that emits a detectable signal. While the preferred reporter dye is a fluorescent dye, any reporter dye that can be attached to a detection reagent, such as an oligonucleotide probe or primer, is suitable according to the invention. In yet another embodiment, the detection reagent may optionally be labeled with a quenching dye, especially when the reagent is used as a self-extinguishing probe, such as the TaqMan probe. The SNP detection reagents described herein may also contain other labels, including, but not limited to, biotin for streptavidin binding, a hapten for antibody binding, and an oligonucleotide for binding to another complementary oligonucleotide.

The present invention also contemplates reagents that do not contain (or are not complementary to) the nucleotide of the SNP to be identified, but which are used to analyze one or more of the SNPs disclosed herein. For example, primers that flank but do not hybridize directly at the site of a target SNP are useful in primer extension reactions in which primers hybridize to a region adjacent to the target SNP position (i.e., within one or more nucleotides of the target SNP site) . During a primer extension reaction, a primer generally cannot extend through the SNP target site if a particular nucleotide (allele) is present at that SNP target site, and the primer extension product can be detected to determine which SNP allele is present at the target site. SNP. For example, specific ddNTPs are typically used in the primer extension reaction to terminate the primer extension, at the time the ddNTP is incorporated into the extension product. Thus, reagents that bind to a nucleic acid molecule in the region adjacent to the SNP site and that are used to analyze the SNP site, even though the associated sequences do not necessarily include the SNP site itself, are also contemplated in accordance with the invention.

The term "FCGR2A" refers to the human FCGR2A gene. This gene encodes the low affinity receptor II-a immunoglobulin Fc region gamma (CD32) and is one member of the immunoglobulin Fc receptor gene family. The protein encoded by this gene is a cell surface receptor found on phagocytic cells such as macrophages and neutrophils and is involved in the process of phagocytosis and clearance of immune complexes. Alternative splicing gives rise to many transcriptional variants.

Preferably, the term "FCGR2A" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 61 of the sequence listing or variant of said nucleotide sequence, and the protein encoded by that nucleic acid, preferably a protein, preferably consisting of the amino acid sequence of SEQ ID NO: 62 sequence listings or variants of the indicated amino acid sequence.

Rs1801274 is a SNP in the FCGR2A gene. The Rs1801274 (C) allele encodes the arginine (R) allele and the (T) allele encodes the histidine (H) variant. This SNP is an intragenic transition followed by a change in codons: CAT, CGT and leads to a missense mutation. SNP is known in the literature under many names, including A519C and R131H. The context sequence looks like this:

TGGGATGGAGAAGGTGGGATCCAAA[C/T]GGGAGAATTTCTGGGATTTTTCCATT

The term "MUC1" refers to the human MUC1 gene. This gene encodes cell surface-associated mucin 1 (MUC1) or polymorphic epithelial mucin (PEM), which is a member of the mucin family and is a membrane-bound glycosylated phosphoprotein. The protein is attached to the apical surface of many epithelia by a transmembrane domain. Behind the transmembrane domain is the SEA domain, which contains a cleavage site to release a large extracellular domain. The protein performs a protective function by binding to pathogens and also functions in cellular signaling.

Preferably, the term "MUC1" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 63 of a sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein comprising, preferably consisting of, the amino acid sequence of SEQ ID NO: 64 sequence listing or variant of the specified amino acid sequence.

Rs4072037 is a SNP in the MUC1 gene. This SNP is an intragenic transition with the following codon change: ACA, ACG and leads to a silent mutation. The context sequence looks like this:

CCCCTAAACCCGCAACAGTTGTTAC[A/G]GGTTCTGGTCATGCAAGCTCTACCC

The term "IL-10" refers to the human IL-10 gene. This gene codes for interleukin-10 (IL-10), also known as human cytokine synthesis inhibitory factor (CSIF), which is an anti-inflammatory cytokine.

Preferably, the term "IL-10" refers to a nucleic acid comprising, preferably consisting of the nucleotide sequence of SEQ ID NO: 65 of the sequence listing or variant of said nucleotide sequence, and the protein encoded by that nucleic acid, preferably a protein containing, preferably consisting of amino acid sequence SEQ ID NO: 66 of a sequence listing or variant of said amino acid sequence.

Rs1800896 is a SNP in the IL-10 gene. This SNP represents an intergenic/unknown intragenic transition. The context sequence looks like this:

CAACACTACTAAGGCTTCTTTGGGA[A/G]GGGGAAGTAGGGATAGGTAAGAGGA

The term "DNMT3A" refers to the human DNMT3A gene. This gene encodes DNA (cytosine-5)-methyltransferase 3A. The protein encoded by this gene is an enzyme that catalyzes the transfer of methyl groups to specific CpG structures in DNA.

Preferably, the term "DNMT3A" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 67 of the sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein containing, preferably consisting of the amino acid sequence SEQ ID NO: 68 of a sequence listing or variant of the indicated amino acid sequence.

Rs1550117 is a SNP in the DNMT3A gene. This SNP is an intragenic transition in the DNMT3A promoter region. The context sequence looks like this:

AATTCCACCAGCACAGCCACTCACT[A/G]TGTGCTCATCTCACTCCTCCAGCAG

The term "SMAD4" refers to the human SMAD4 gene. This gene encodes the 4th homologue of Mothers against decapentaplegic. The protein encoded by this gene is involved in cellular signaling and belongs to the family of Darwin proteins that modulate members of the TGFβ protein superfamily. The protein binds receptor-regulated SMADs such as SMAD1 and SMAD2 and forms a complex that binds to DNA and serves as a transcription factor. It is the only known co-SMAD mammal.

Preferably, the term "SMAD4" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 69 of a sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein comprising, preferably consisting of, the amino acid sequence of SEQ ID NO: 70 of the sequence listing or variant of the specified amino acid sequence.

Rs12456284 is a SNP in the SMAD4 gene. This SNP is an intragenic transition in the 3'-UTR. The context sequence looks like this:

AGGTCCAGAGCCAGTGTTCTTGTTC[A/G]ACCTGAAAGTAATGGCTCTGGGTTG

The term "EGF" refers to the human EGF gene. This gene codes for epidermal growth factor. EGF is a growth factor that stimulates cell growth, proliferation and differentiation by binding to its EGFR receptor.

Preferably, the term "EGF" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 71 of the sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein comprising, preferably consisting of the amino acid sequence of SEQ ID NO: 72 sequence listing or variant of the indicated amino acid sequence.

Rs4444903 is a SNP in the EGF gene. This SNP is an intragenic transition in the 5'-UTR. The context sequence looks like this:

CTTTCAGCCCCAATCCAAGGGTTGT[A/G]GCTGGAACTTTCCATCAGTTCTTCC

The term "CDH1" refers to the human CDH1 gene. This gene codes for cadherin-1, also known as CAM 120/80 or epithelial cadherin (E-cadherin) or uvomorulin. The protein is a classic member of the cadherin superfamily. The protein is a calcium-dependent intercellular adhesion glycoprotein consisting of five extracellular cadherin repeats, a transmembrane region, and a highly conserved cytoplasmic tail. Loss of function is thought to promote cancer progression by enhancing proliferation, invasion and/or metastasis.

Preferably, the term "CDH1" refers to a nucleic acid comprising, preferably consisting of the nucleotide sequence of SEQ ID NO: 73 of the sequence listing or variant of said nucleotide sequence, and the protein encoded by that nucleic acid, preferably a protein containing, preferably consisting of the amino acid sequence SEQ ID NO: 74 sequence listings or variants of the indicated amino acid sequence.

Rs16260 is a SNP in the CDH1 gene. This SNP is an intragenic transition located in the promoter region of the CDH1 gene. The context sequence looks like this:

CTAGCAACTCCAGGCTAGAGGGTCA[A/C]CGCGTCTATGCGAGGCCGGGTGGGC

The term "ERCC1" refers to the human ERCC1 gene. This gene encodes the ERCC-1 repair protein. The function of the ERCC1 protein is mainly to excision repair of damaged DNA.

Preferably, the term "ERCC1" refers to a nucleic acid comprising, preferably consisting of the nucleotide sequence of SEQ ID NO: 75 of the sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein containing, preferably consisting of the amino acid sequence SEQ ID NO: 76 of a sequence listing or variant of the indicated amino acid sequence.

Rs11615 is a SNP in the ERCC1 gene. This SNP is a silent intragenic transition. The context sequence looks like this:

ATCCCGTACTGAAGTTCGTGCGCAA[C/T]GTGCCCTGGGAATTTGGCGACGTAA

The term "FCGR3A" refers to the human FCGR3A gene. This gene encodes a low affinity immunoglobulin gamma Fc receptor region, type III-A. The protein encoded by this gene is part of a cluster of surface molecules of differentiating cells.

Preferably, the term "FCGR3A" refers to a nucleic acid comprising, preferably consisting of, the nucleotide sequence of SEQ ID NO: 77 of a sequence listing or variant of said nucleotide sequence, and a protein encoded by that nucleic acid, preferably a protein comprising, preferably consisting of, the amino acid sequence of SEQ ID NO: 78 of the sequence listing or variant of the specified amino acid sequence.

Rs396991 is a SNP in the FCGR3A gene. This SNP is an intragenic transversion with the following codon change: GTT, TTT and results in a missense mutation. Rs396991 (T) encodes the phenylalanine (F) allele, with the (G) allele encoding the valine variant (v). The context sequence looks like this:

CGGCTCCTACTTCTGCAGGGGGCTT[G/T]TTGGGAGTAAAAATGTGTCTTCAGA

Claudins are a family of proteins that are the most important components of tight junctions, where they establish a paracellular barrier that controls the flow of molecules in the intercellular space between epithelial cells. Claudins are transmembrane proteins that cross the membrane 4 times, with the N- and C-terminus located in the cytoplasm. The first extracellular loop or domain consists of an average of 53 amino acids, and the second extracellular loop or domain consists of about 24 amino acids. Cell surface proteins of the claudin family, such as CLDN18.2, are expressed in tumors of various origins and are particularly suitable as target structures in the case of antibody-mediated cancer immunotherapy due to their selective expression (lack of expression in toxicity-relevant normal tissue) and localization in the plasma membrane.

As used herein, the term "CLDN" means claudin and includes CLDN18.2. Preferably the claudin is human claudin.

The term "CLDN18" refers to claudin 18 and includes any variants including claudin 18 splicing variant 1 (claudin 18.1 (CLDN18.1)) and claudin 18 splicing variant 2 (claudin 18.2 (CLDN18.2)).

The term "CLDN18.2" preferably refers to human CLDN18.2 and in particular to a protein containing, preferably consisting of, the amino acid sequence according to SEQ ID NO: 1 of the sequence listing or a variant of said amino acid sequence. The first extracellular loop or CLDN18.2 domain preferably contains amino acids 27 to 81, more preferably amino acids 29-78 of the amino acid sequence shown in SEQ ID NO: 1. The second extracellular loop or CLDN18.2 domain preferably contains amino acids 140-180 of the amino acid sequence, shown in SEQ ID NO: 1. Said first and second extracellular loops or domains preferably form the extracellular portion of CLDN18.2.

CLDN18.2 is selectively expressed in normal tissues in differentiated epithelial cells of the gastric mucosa. CLDN18.2 is expressed in malignancies of various origins such as pancreatic carcinoma, esophageal carcinoma, gastric carcinoma, bronchial carcinoma, breast carcinoma, and ENT tumors. CLDN18.2 is a valuable target for the prevention and/or treatment of primary tumors such as gastric cancer, esophageal cancer, pancreatic cancer, lung cancer such as non-small cell lung cancer (NSCLC), ovarian cancer, colon cancer, liver cancer, cancer head and neck, gallbladder cancer and metastases, in particular gastric cancer metastases such as Krukenberg tumors, peritoneal metastases and lymph node metastases.

The term "CLDN18.1" preferably refers to human CLDN18.1 and in particular to a protein containing, preferably consisting of, the amino acid sequence according to SEQ ID NO: 2 of the sequence listing or variant of said amino acid sequence.

"Prognosis", as used herein, refers to the prediction of outcome and, in particular, the likelihood of progression-free survival (PFS) or disease-free survival (disease-free survival, DFS). Survival is usually calculated as the average number of months (or years) during which 50% of patients survive, or the percentage of patients who are still alive after 1, 5, 15 and 20 years. Prognosis is important for treatment decisions because patients with a good prognosis are usually offered less invasive treatments, while those with a poor prognosis are usually offered more aggressive treatments, such as more extensive chemotherapy drugs.

"Prediction", as used herein, refers to providing information about the likely response of a disease to a particular therapeutic intervention.

The phrase "indicates risk" refers to indicating a certain degree of likelihood or plausibility. The phrase "indicates risk reduction" refers to a low degree of likelihood or plausibility. The phrase "indicates an increased risk" refers to a certain, greater, or high degree of likelihood or likelihood.

If the event "indicates a reduced risk that the cancer patient is not a responder to antibody treatment", the event indicates that the cancer patient is a responder to antibody treatment, i.e. the patient is likely to be a treatment responder antibody and, optionally, it is more likely that the patient is a responder to antibody treatment than that the patient is not a responder to antibody treatment.

If an event "indicates an increased risk that the cancer patient is not a responder to antibody treatment", the event indicates that the cancer patient is not a responder to antibody treatment, i.e. the patient is likely not a responder antibody treatment, and optionally, it is more likely that the patient is not a responder to antibody treatment than that the patient is a responder to antibody treatment.

If an event "indicates a reduced risk of an adverse clinical outcome", said event indicates a favorable clinical outcome, that is, it is likely that there will be a favorable clinical outcome, and it may be more likely that there will be a favorable clinical outcome than that there will be an unfavorable clinical outcome .

If an event "indicates an increased risk of an adverse clinical outcome", said event is indicative of an adverse clinical outcome, i.e. it is likely to have an adverse clinical outcome, and possibly more likely to have an adverse clinical outcome than a favorable clinical outcome.

If an event "indicates a reduced risk that the cancer patient will not experience progression-free survival", said event indicates that the cancer patient will experience progression-free survival, i.e. it is likely that the patient will experience progression-free survival. progression-free survival and, optionally, more likely to experience progression-free survival than that the patient does not experience progression-free survival.

If an event "indicates an increased risk that the cancer patient will not experience progression free survival", then said event indicates that the cancer patient will not experience progression free survival, i.e. it is likely that the patient will not experience with progression free survival, and optionally it is more likely that the patient will not experience progression free survival than that the patient will experience progression free survival.

Used in this document, the term "sample" refers to any material that is obtained from the subject and which can be used for analytical purposes, in particular to determine the genotype of one or more single nucleotide polymorphisms. In some embodiments, the samples described herein can be or can be obtained from any tissues, cells and/or cells in biological fluids, for example, from a mammal or human being to be tested. The sample may be isolated from a patient, for example from a human body. The sample may be a fractionated and/or purified sample. For example, samples covered by the present invention may be or may be obtained from tissue samples (e.g., a section or explant), single-cell samples, cell colony samples, cell culture samples, blood (e.g., whole blood or a blood fraction such as blood cells, serum or plasma), urine samples or samples from other peripheral sources. In one particularly preferred embodiment, the sample is a tissue sample (eg, a biopsy of an object suspected of having malignant tumor tissue). For example, the sample may be a tumor biopsy. The sample may be obtained from the patient prior to initiation of treatment, during treatment, and/or after treatment, for example before, during, or after administration of anticancer therapy.

Samples can be used to obtain nucleic acid (including DNA and/or RNA), protein or membrane extracts from any body fluids (such as blood, serum, plasma, urine, saliva, sputum, gastric juices, semen, tears, sweat and etc.), skin, hair, cells (especially germ cells), biopsies, buccal swabs or tissue or tumor specimens.

The present invention also relates to a kit containing tools, such as reagents for determining the genotype of one or more single nucleotide polymorphisms, as described in this document. In the context of the present invention, the term "composite set (shortly: set)" refers to any combination of at least some of the number of components designated herein, which are combined, coexist spatially, in a functional unit, and which may contain additional components. For example, the kit may include preselected primers or probes specific for nucleic acid sequences containing one or more single nucleotide polymorphisms whose genotype is to be determined. The kit may also contain enzymes suitable for nucleic acid amplification (eg polymerases such as Taq) and deoxynucleotides and buffers necessary for the amplification reaction mixture. The kit may also contain probes specific for one or more single nucleotide polymorphisms. In some embodiments, said agents are detectably labelled.

The kit according to the invention may contain (i) a container and/or (ii) a data carrier. Said container may be filled with one or more of the aforementioned agents or reagents. Said storage medium may be a non-electronic storage medium, such as a graphic storage medium such as an information brochure, information sheet, barcode or access code, or an electronic storage medium such as a floppy disk, compact disc (CD), digital versatile disk (DVD). ), microchip or other electronic media based on semiconductors. The access code may allow access to a database, such as an internet database, a centralized database, or a decentralized database. Said data carrier may contain instructions for providing analysis of the results obtained with said kit, and in particular for using the kit in the methods of the invention.

Additionally or alternatively, said kit may contain commercially and user-desirable materials, including buffer(s), reagent(s), and/or diluent(s).

Based on the results obtained (ie, based on the genotype for one or more single nucleotide polymorphisms), the clinician may select an antitumor therapy to which the patient is predicted to be responsive, in particular antibody therapy. Preferably, an anticancer therapy to which the patient is predicted to be refractory is not administered to the patient.

Based on the fact that the patient is predicted to be refractory to antibody therapy, in particular antibody therapy, which acts by recruiting the patient's immune system to destroy tumor cells, the physician may choose to use anticancer therapy that is different from antibody therapy, in particular, antibody therapy, which acts by recruiting the patient's immune system to destroy tumor cells. In particular, the doctor may choose chemotherapy.

Based on the fact that the patient is predicted to be responsive to antibody therapy, in particular antibody therapy, acting by recruiting the patient's immune system to kill tumor cells, the clinician may choose to treat with an antibody, in particular, antibody therapy, acting by recruiting the patient's immune system to kill tumor cells, optionally in combination with chemotherapy.

The term "therapeutic treatment", in particular in connection with the treatment of cancer, as used herein, refers to any treatment that is aimed at improving the health status and/or prolonging (increasing) life expectancy of a patient. Said treatment may eliminate a cancer, reduce the size or number of tumors in a patient, stop or slow the progression of a cancer in a patient, inhibit or slow the development of a new cancer in a patient, reduce the frequency or severity of symptoms in a patient, and/or reduce recurrence in a patient who currently suffering from or who previously had a malignancy. The (therapeutic) effect on the cancer may be selected from the group consisting of surgery, chemotherapy, radiation therapy, and targeted therapy. One particularly preferred method of treatment according to the invention is the treatment of cancer using therapeutic monoclonal antibodies against tumor antigens, such as CLDN18.2, expressed on target cells.

Adjuvant therapy is a treatment that is given in addition to primary, main or initial treatment. The surgical intervention and complex treatment regimens used in the treatment of malignancy lead to the term being used primarily to describe the adjuvant treatment of malignancy. An example of adjuvant therapy is an adjunctive treatment usually given after surgery when all detected disease has been removed, but which retains a statistical risk of recurrence due to undiagnosed disease.

Terms such as "responsive" or "responder" refer in a therapeutic setting to the fact that a patient has a therapeutic benefit from a given treatment and, in particular, to seeing the reduction, prevention, or elimination of a disease, including shortening the duration of the disease, stopping or slowing the progression or worsening of a disease, inhibiting or slowing the development of a new disease and/or relapses, preventing or delaying the onset of a disease or its symptoms, reducing the frequency or severity of symptoms in a patient who is currently suffering from or has previously had a disease, and/or to prolong the life of the patient. In particular, they refer to the observation of a reduction in tumor mass, or an increase in tumor-free period, disease-free period, or overall survival period.

Terms such as "non-responsive" or "non-responder" refer in a therapeutic setting to the fact that a patient does not benefit from a given treatment and, in particular, there is no amelioration, prevention or elimination of the disease, i.e. the patient is resistant to treatment. .

A complete response is defined as the absence of any residual disease, such as malignancy, and is usually assessed by pathological analysis of extracted tissue samples. The term "complete pathological response" (pCR) is often used in this context. In particular, pCR is defined as the absence of any residual invasive tumor cells in the original tumor bed. However, the definition of pCR may vary between different classification systems. Pathological complete response has been shown to be a predictive factor for overall better survival, but also for disease-free survival and disease-free survival.

Disease-free survival is defined as the time from randomization to any first of a number of relapses or recurrences of the disease, a second malignancy, or death.

Progression-free survival (PFS) is a type of survival that measures the length of time during and after treatment or treatment in which the disease being treated (usually cancer) does not get worse. The term is sometimes used as a proxy for examining the health of a person with a disease to try to determine how well a new treatment works, and is often used as a clinical endpoint in randomized controlled trials for the treatment of cancer.

According to the invention, the term "cancer patient experiencing progression-free survival" refers to a cancer patient who has no progression of the disease over a long period of time, in particular compared to the average period in patients and/or compared to the period in patients who are non-responders to this treatment regimen. Preferably, said long period of time is at least 4, preferably at least 5, more preferably at least 6 months, such as at least 7 months or at least 8 months, wherein said the time period starts, for example, from the first administration of the treatment.

The term "clinical outcome" is defined as the clinical outcome of a disease, such as reduction or relief of symptoms, particularly after treatment.

The term "relapse" in relation to a malignant tumor includes the appearance of tumor cells in the same place and in the organ of origin of the disease, distant metastases that may appear many years after the initial diagnosis and antitumor therapy, or in local events such as infiltration of tumor cells into regional lymph nodes.

The terms "individual" and "object" are used interchangeably herein. They refer to humans, non-human primates, or other mammals (e.g., mice, rats, rabbits, dogs, cats, cows, pigs, sheep, horses, or primates) that may be affected by or susceptible to a disease or disorder (e.g., malignant neoplasm), but may or may not have a disease or disorder. In many embodiments, the individual is a human. Unless otherwise indicated, the terms "individual" and "object" do not designate a specific age and, therefore, cover adults, the elderly, children, and newborns. In preferred embodiments of the present invention, the "individual" or "subject" is a "patient". The term "patient" means in accordance with the invention an object for treatment, in particular a sick object.

In one particularly preferred embodiment, the method of the invention is performed on a patient who has already been diagnosed as suffering from a malignant neoplasm.

"Target cell" means any unwanted cell, such as a cancer cell. In preferred embodiments, the target cell expresses CLDN18.2.

In the context of the present invention, terms such as "protection", "prevention" or "prophylaxis" refer to preventing the occurrence and/or spread of a disease in a subject and, in particular, to minimizing the likelihood that the subject will develop a disease or to delay the development of the disease. For example, a subject at risk of developing cancer would be a candidate for cancer prevention therapy.

By "threatened" is meant an entity that is identified as having a higher than normal chance of developing a disease, in particular cancer, compared to the general population. In addition, an object that has or currently has a disease, in particular cancer, is an object that is at increased risk of developing the disease because such an object may continue to develop the disease. Subjects that currently have or have had cancer also have an increased risk of developing cancer metastases.

Used in this document, the term "combination" in the context of the introduction of therapy refers to the use of more than one therapeutic agent or therapeutic agent. The use of the term "in combination" does not limit the order in which the therapeutic agents or therapeutic agents are administered to the subject. Therapeutic agents or therapeutic agents can be administered before, simultaneously with, or after administration of the second therapy or therapeutic agent to the subject. Preferably, the therapies or therapeutic agents are administered to the subject in sequence, amount, and/or over a time interval so that the therapeutics or therapeutic agents can act together. In a specific embodiment, the therapeutic agents or therapeutic agents are administered to the subject in sequence, amount and/or over a time interval such that they provide an increased benefit than if they were otherwise administered, in particular independently of each other. Preferably, the increased benefit is a synergistic effect.

The term "disease" refers to an abnormal condition that affects the human body. The disease is often interpreted as a medical condition associated with specific symptoms and symptoms. The disease may be caused by factors originating from an external source, such as an infectious disease, or may be caused by internal dysfunctions, such as autoimmune diseases. In humans, "disease" is often used more broadly to refer to any condition that causes pain, dysfunction, distress, social problems, or death for individual sufferers or similar problems for those in contact with the person. In this broader sense, disease sometimes includes injuries, disabilities, disorders, syndromes, infections, isolated symptoms, deviant behavior, and atypical variations in structure and function, while in other contexts and for other purposes they may be considered distinct categories. Illnesses usually affect people not only physically, but also emotionally, as acquiring and living with many diseases can change the outlook on an individual's life and personality. According to the invention, the term "disease" includes a malignant neoplasm, in particular those forms of malignant neoplasms that are described in this document. Any reference herein to cancer or specific forms of cancer also includes cancer metastases. In a preferred embodiment, the disease to be treated according to the present application comprises cells expressing a tumor antigen such as CLDN18.2.

"Disease associated with cells expressing a tumor antigen" means in accordance with the invention that a tumor antigen, such as CLDN18.2, is expressed in cells of a diseased tissue or organ. In one embodiment, the expression of the tumor antigen in the cells of the diseased tissue or organ is increased compared to the state in a healthy tissue or organ. Increase refers to an increase of at least 10%, in particular at least 20%, at least 50%, at least 100%, at least 200%, at least at least 500%, at least 1000%, at least 10000% or even more. In one embodiment, the expression is found only in the diseased tissue, while the expression in the corresponding healthy tissue is repressed. According to the invention, diseases associated with cells expressing a tumor antigen include oncological diseases. In addition, according to the invention, cancers are preferably diseases in which cancer cells express a tumor antigen.

The terms "cancer" or "malignant neoplasm" refer to the physiological state of the individual or describe his physiological state, which is usually characterized by unregulated cell growth. Examples of malignancies include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. Specifically, examples of such malignancies include malignancies of the bone, blood, lung, liver, pancreas, skin, head or neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, gastric cancer, cancer colon cancer, breast cancer, prostate cancer, uterine cancer, genital and reproductive organ carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, soft tissue sarcoma, cancer bladder, kidney cancer, renal cell carcinoma, renal pelvis carcinoma, central nervous system (CNS) neoplasia, neuroectodermal cancer, spinal axis tumors, glioma, meningioma and pituitary adenoma. The term "malignant neoplasm" in accordance with the invention also includes metastases of malignant neoplasms. Preferably, the "cancer" is characterized by cells expressing a tumor antigen such as CLDN18.2 and the cancer cell expresses such tumor antigen. A cell expressing a tumor antigen such as CLDN18.2 is preferably a cancer cell, preferably from the cancers described herein.

According to the invention, the term "tumor" or "neoplastic disease" refers to the abnormal growth of cells (termed neoplastic cells, tumor cells or tumor cells), preferably forming a tumor or tumor. By "tumor cell" is meant an abnormal cell that grows by rapid, uncontrolled cell proliferation and continues to grow after the stimuli that initiated the new growth have ceased. Tumors exhibit partial or complete lack of structural organization and functional coordination with normal tissue and usually form a distinct mass of tissue that may be either benign, premalignant, or malignant.

In one embodiment, the cancer according to the invention comprises cancer cells expressing a tumor antigen, such as CLDN18.2. In one embodiment, the cancer is a tumor antigen such as CLDN18.2 positive. In one embodiment, expression of a tumor antigen, such as CLDN18.2, is on the cell surface. In one embodiment, at least 50%, preferably 60%, 70%, 80%, or 90% of the cancer cells are tumor antigen positive, e.g., CLDN18.2 positive and/or at least 40%, preferably, at least 50% of the cancer cells are positive for surface expression of a tumor antigen, such as CLDN18.2. In one embodiment, at least 95% or at least 98% of cancer cells are positive for a tumor antigen, such as CLDN18.2. In one embodiment, at least 60%, at least 70%, at least 80%, or at least 90% of cancer cells are positive for surface expression of a tumor antigen, such as CLDN18.2.

In one embodiment, the cancer comprising cancer cells expressing CLDN18.2 or CLDN18.2 positive cancer is selected from the group consisting of gastric cancer, esophageal cancer, pancreatic cancer, lung cancer such as non-small cell lung cancer (NSCLC), ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder cancer and metastases, in particular gastric cancer metastases such as Krukenberg tumors, peritoneal metastasis and lymph node metastases. In one embodiment, the cancer is an adenocarcinoma, in particular a progressive adenocarcinoma. Particularly preferred cancers are adenocarcinomas of the stomach, esophagus, pancreatic ducts, bile ducts, lung and ovary. In one embodiment, the cancer is selected from the group consisting of gastric cancer, cancer of the esophagus, in particular the lower esophagus, cancer of the esophagogastric junction, and gastroesophageal cancer. In a particularly preferred embodiment, the cancer is gastroesophageal cancer, such as metastatic, refractory, or recurrent progressive gastroesophageal cancer. In one embodiment, the CLDN18.2 positive tumor is a tumor of the above types of malignancies.

Embodiments involving a CLDN18.2 positive tumor or cancer cells expressing CLDN18.2 preferably include the use of an antibody having the ability to bind to CLDN18.2. In one embodiment, the antibody having the ability to bind to CLDN18.2 is a monoclonal, chimeric, or humanized antibody or antibody fragment.

According to the invention, "carcinoma" is a malignant tumor derived from epithelial cells. This group represents the most common types of malignant neoplasms, including common forms of breast, prostate, lung and colon cancer.

"Adenocarcinoma" is a malignant neoplasm that occurs in glandular tissue. This tissue is also part of a broader category of tissue known as epithelial tissue. Epithelial tissue includes the skin, glands, and many other tissues that line the cavities and organs of the body. The epithelium is derived embryologically from the ectoderm, endoderm and mesoderm. To be classified as an adenocarcinoma, the cells do not have to be part of a gland if they have secretory properties. This form of carcinoma can occur in some higher mammals, including humans. Well-differentiated adenocarcinomas tend to resemble the glandular tissue from which they originate, while poorly differentiated ones may not. By staining the biopsy cells, the pathologist will determine if the tumor is an adenocarcinoma or another type of cancer. Adenocarcinomas can occur in many body tissues due to the ubiquitous nature of glands in the body. Although each gland cannot secrete the same substance, as long as it functions exocrine in the cell, it is considered glandular, so its malignant form is called adenocarcinoma. Malignant adenocarcinomas invade other tissues and often metastasize if given enough time. Ovarian adenocarcinoma is the most common type of ovarian carcinoma. It includes serous and mucinous adenocarcinoma, renal cell carcinoma, and endometrioid adenocarcinoma.

By "metastasis" is meant the spread of cancer cells from their original site to another part of the body. The formation of metastases is a very complex process and depends on the detachment of malignant cells from the primary tumor, invasion of the extracellular matrix, penetration through the endothelial basement membranes into the body cavity and blood vessels, and then, after transport by blood, infiltration of target organs. Finally, the growth of a new tumor at the target site depends on angiogenesis. Tumor metastases often occur even after removal of the primary tumor because tumor cells or components may remain and develop metastatic potential. In one embodiment, the term "metastasis" according to the invention refers to "distant metastasis", which refers to metastasis remote from the primary tumor and the regional lymph node system. In one embodiment, the term "metastasis" in accordance with the invention refers to metastasis to the lymph nodes. One particular form of metastasis that is treatable using the therapy of the invention is metastasis originating from gastric cancer as the primary site. In preferred embodiments, such gastric cancer metastasis is Krukenberg tumors, peritoneal metastasis, and/or lymph node metastasis.

A refractory malignancy is a malignant tumor for which a specific treatment is ineffective, or which initially does not respond to treatment, or becomes resistant over time. The terms "refractory", "non-responsive" or "resistant" are used interchangeably herein.

Krukenberg's tumor is an uncommon metastatic ovarian tumor accounting for 1% to 2% of all ovarian tumors. The prognosis of Krukenberg tumor is still very poor and there is no cure for Krukenberg tumors. Krukenberg's tumor is a metastatic ring cell adenocarcinoma of the ovary. The stomach is the main site in most cases of Krukenberg tumors (70%). Colon, appendix, and breast carcinomas (predominantly invasive lobular carcinoma) are the most common primary sites. Rare cases of Krukenberg tumor originating from carcinomas of the gallbladder, biliary tract, pancreas, small intestine, ampulla of Vater, cervix, and bladder/urachus have been reported.

The term "surgery", as used in this document, includes the removal of tumors during surgery. It is a common treatment for malignant neoplasms. The surgeon may remove tumors using local excision.

As used herein, the term "chemotherapy" refers to the use of chemotherapeutic agents or combinations of chemotherapeutic agents, preferably to stop the growth of cancer cells, either by killing the cells or stopping them from dividing. When chemotherapy is taken by mouth or injected into a vein or muscle, the drugs enter the bloodstream and can enter cancer cells throughout the body (systemic chemotherapy). When chemotherapy is placed directly into the cerebrospinal fluid, an organ, or a body cavity such as the abdomen, the drugs primarily target cancer cells in those areas (regional chemotherapy).

The chemotherapeutic agents of the invention include cytostatic compounds and cytotoxic compounds. Traditional chemotherapeutic agents work by killing cells that are rapidly dividing, which is one of the main properties of most cancer cells. This means that chemotherapy also harms cells that divide rapidly under normal circumstances, such as cells in the bone marrow, digestive tract, and hair follicles. This leads to the most common side effects of chemotherapy. According to the invention, the term "chemotherapy" preferably does not include antibodies that target proteins that are abnormally expressed in cancer cells (tumor antigens) and act by recruiting the patient's immune system to kill tumor cells. However, antibodies that target proteins that are abnormally expressed in cancer cells (tumor antigens) and act through a therapeutic moiety or antibody-conjugated agent may be considered a form of chemotherapy. However, in the strict sense, the term "chemotherapy" in accordance with the invention does not include targeted therapy.

According to the invention, the term "chemotherapeutic agent" includes taxanes, platinum compounds, nucleoside analogs, camptothecin analogs, anthracyclines, etoposide, bleomycin, vinorelbine, cyclophosphamide, and combinations thereof. According to the invention, a reference to a chemotherapeutic agent includes any prodrug, such as an ester, salt, or derivative, such as a conjugate of said agent. Examples are conjugates of said agent with a carrier, for example protein-bound paclitaxel, such as albumin-bound paclitaxel. Preferably, the salts of said agent are pharmaceutically acceptable.

Taxanes are a class of diterpene compounds that were first obtained from natural sources such as plants of the genus Taxus, but some have been artificially synthesized. The main mechanism of action of taxane-class drugs is the disruption of microtubule function, which interferes with the process of cell division. Taxanes include docetaxel (Taxotere) and paclitaxel (Taxol).

According to the invention, the term "docetaxel" refers to a compound having the following formula:

In particular, the term "docetaxel" refers to the compound 1,7β,10β-trihydroxy-9-oxo-5β,20-epoxytax-11-en-2α,4,13α-triyl-4-acetate 2-benzoate 13-{( 2R,3S)-3-[(tert-butoxycarbonyl)amino]-2-hydroxy-3-phenylpropanoate}.

According to the invention, the term "paclitaxel" refers to a compound having the following formula:

In particular, the term "paclitaxel" refers to the compound (2α, 4α, 5β, 7β, 10β, 13α)-4,10-bis-(acetyloxy)-13-{[(2R,3S)-3-(benzoylamino)) -2-hydroxy-3-phenylpropanoyl]oxy}-1,7-dihydroxy-9-oxo-5,20-epoxytax-11-en-2-ylbenzoate.

In accordance with the invention, the term "platinum compound" refers to compounds containing platinum in their structure, such as platinum complexes, and includes compounds such as cisplatin, carboplatin and oxaliplatin.

The term "cisplatin" or "cisplatin" refers to a cis-diammedichloroplatinum (ii) (CDDP) compound of the following formula:

The term "carboplatin" refers to a cis-diamine (1,1-cyclobutanedicarboxylate) platinum (ii) compound of the following formula:

The term "oxaliplatin" refers to a compound which is a platinum compound which is complexed with a diaminocyclohexane-bearing ligand of the following formula:

In particular, the term "oxaliplatin" refers to the compound [(1R, 2R)-cyclohexane-1,2-diamine] (ethanedioato-O,O') platinum (ii). Oxaliplatin for injection is also sold under the brand name Eloxatin.

The term "nucleoside analog" refers to a structural nucleoside analog, a category that includes both purine analogs and pyrimidine analogs.

The term "gemcitabine" is a compound that is a nucleoside analog of the following formula:

In particular, this term refers to the compound 4-amino-1- (2-deoxy-2,2-difluoro-β-D-erythro-pentofuranosyl) pyrimidin-2 (1H)-one or 4-amino-1- [ ( 2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-1,2-dihydropyrimidin-2-one.

The term "nucleoside analog" includes fluoropyrimidine derivatives such as fluorouracil and its prodrugs. The term "fluorouracil" or "5-fluorouracil" (5-FU or f5U) (sold under the trade names Adrucil, Carac, Efudix, Efudex, and Fluoroplex) is a compound that is a pyrimidine analog of the following formula:

In particular, this term refers to the compound 5-fluoro-1H-pyrimidine-2,4-dione.

The term "capecitabine" (Xeloda, Roche) refers to a chemotherapeutic agent that is a prodrug that is converted to 5-FU in tissues. Capecitabine, which can be administered orally, has the following formula:

In particular, this term refers to the compound pentyl[1-(3,4-dihydroxy-5-methyltetrahydrofuran-2-yl)-5-fluoro-2-oxo-1H-pyrimidin-4-yl] carbamate.

The term "folinic acid" or "leucovorin" refers to a compound useful in synergistic combination with the chemotherapeutic agent 5-fluorouracil. Thus, if reference is made herein to the administration of 5-fluorouracil or a prodrug, said administration in one embodiment may include administration in combination with folinic acid. Folinic acid has the following formula:

In particular, this term refers to the compound (2S)-2-{[4-[(2-amino-5-formyl-4-oxo-5,6,7,8-tetrahydro-1H-pteridin-6-yl) )methylamino]benzoyl]amino}pentanedioic acid.

According to the invention, the term "analogue of camptothecin" refers to derivatives of the compound camptothecin (CPT; (S)-4-ethyl-4-hydroxy-1H-pyrano[3',4':6,7] indolizino[1,2-b ]quinoline-3,14-(4H,12H)-dione). Preferably, the term "camptothecin analog" refers to compounds comprising the following structure:

According to the invention, the preferred analogues of camptothecin are inhibitors of DNA topoisomerase I (topo I). Preferred analogues of camptothecin in accordance with the invention are irinotecan and topotecan.

Irinotecan is a drug that prevents DNA degradation by inhibiting topoisomerase I. In chemical terms, it is a semi-synthetic analogue of the natural alkaloid camptothecin, having the following formula:

In particular, the term "irinotecan" refers to the compound (S)-4,11-diethyl-3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo-1H-pyrano[3',4': 6,7]-indolizino-[1,2-b]quinolin-9-yl-[1,4'-bipiperidine]1'-carboxylate.

Topotecan is a topoisomerase inhibitor of the formula:

In particular, the term "topotecan" refers to the compound (S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano [3', 4': 6,7] indolizino [1 ,2-b] quinoline-3,14 (4H, 12H)-dione.

Anthracyclines are a class of drugs commonly used in cancer chemotherapy that are also antibiotics. Structurally, all anthracyclines share the 4-ring structure of 7,8,9,10-tetrahydrotetracen-5,12-quinone and generally require glycosylation at specific sites.

Anthracyclines preferably result in one or more of the following mechanisms of action: 1. Inhibition of DNA and RNA synthesis by intercalation between DNA/RNA chain base pairs, which prevents the replication of rapidly growing cancer cells. 2. Inhibition of the topoisomerase II enzyme, preventing the supercoiled DNA from relaxing and thus blocking DNA transcription and replication. 3. Creation of iron-mediated oxygen free radicals that damage DNA and cell membranes.

According to the invention, the term "anthracycline" preferably refers to an agent, preferably an antitumor agent, for inducing apoptosis, preferably by inhibiting DNA rebinding at topoisomerase II.

Examples of anathracyclines and anthracycline analogs include, but are not limited to, daunorubicin (daunomycin), doxorubicin (adriamycin), epirubicin, idarubicin, rhodomycin, pyrarubicin, valrubicin, N-trifluoroacetyl doxorubicin-14-valerate, aclacinomycin, morpholinodoxorubicin (morpholino-DOX) , cyanomorpholino-doxorubicin (cyanomorpholino-DOX), 2-pyrrolino-doxorubicin (2-PDOX), 5-iminodunomycin, mitoxantrone, and aclacinomycin A (aclarubicin). Mitoxantrone is a member of the anthracenedione class of compounds, which are analogues of anthracyclines that do not have the sugar moiety of the anthracyclines but retain a planar polycyclic aromatic ring structure that allows for DNA intercalation.

Specifically considered as an anthracycline in the context of the present invention is epirubicin. Epirubicin is an anthracycline drug that has the following formula:

and is sold under the trade name Ellence in the US and Pharmorubicin or Epirubicin Ebewe elsewhere. In particular, the term "epirubicin" refers to the compound (8R, 10S)-10-[(2S, 4S, 5R, 6S)-4-amino-5-hydroxy-6-methyl-oxan-2-yl] oxy-6 ,11-dihydroxy-8-(2-hydroxyacetyl)-1-methoxy-8-methyl-9,10-dihydro-7H-tetracen-5,12-dione. Epirubicin is preferred over doxorubicin and is the most popular anthracycline in some chemotherapy regimens because it appears to cause fewer side effects.

The term "etoposide" refers to a semi-synthetic podophyllotoxin derivative exhibiting antitumor activity. Etoposide inhibits DNA synthesis by complexing with topoisomerase II and DNA. This complex induces breaks in double-stranded DNA and prevents repair by topoisomerase II binding. Accumulated DNA breaks prevent penetration into the mitotic phase of cell division and lead to cell death. Etoposide has the following formula:

In particular, this term refers to the compound 4'-demethyl-epipodophyllotoxin 9-[4,6-O-(R)-ethylidene-beta-D-glucopyranoside], 4'-(dihydrogen phosphate).

The term "bleomycin" refers to a glycopeptide antibiotic produced by the bacterium Streptomyces verticillus. When used as an anticancer agent, it works by causing breaks in DNA. Bleomycin preferably includes a compound having the following formula:

The term "vinorelbine" refers to an antimitotic chemotherapy drug that is a semi-synthetic vinca alkaloid and is provided as a treatment for certain types of cancer, including breast cancer and non-small cell lung cancer. Vinorelbine preferably includes a compound having the following formula:

Cyclophosphamide is a mustard alkylating agent from the group of oxazophorins. The main use of cyclophosphamide is associated with other chemotherapeutic agents in the treatment of certain forms of malignancy. Cyclophosphamide preferably includes a compound having the following formula:

In the context of the present invention, the term "radiotherapy" refers to the use of high-energy x-rays or other types of radiation to kill cancer cells or to prevent their growth. There are two types of radiation therapy. External beam radiation therapy uses a machine outside the body to deliver radiation to a cancerous tumor. Internal radiation therapy uses radioactive material sealed in needles, seeds, wires, or catheters that are placed directly into or near the cancerous tumor. The method of radiation therapy depends on the type and stage of treatment of malignant neoplasms.

In accordance with the invention, the term "targeted therapy" refers to any therapy that can be used to target predominantly diseased cells, such as cancer cells, and which does not target or targets unaffected cells to a lesser extent. Targeting diseased cells preferably results in killing and/or impairing the proliferation or viability of diseased cells. Such therapy includes: i) antibodies, antibody fragments, and proteins that are either uncoated or conjugated to the therapeutic moiety that target specific cell surface targets on diseased cells, such as tumor antigens, such as CLDN18.2 (e.g., antibodies or conjugates antibodies against CLDN18.2 as described herein) or ii) small molecules that disrupt the proliferation or viability of affected cells. In a specific embodiment, the agent binds to an antigen that is expressed at a higher level on diseased than on normal stem cells. In a particular embodiment, the agent binds specifically to a tumor antigen. Conventional chemotherapy or radiation therapy is not considered "targeted therapy" despite the fact that they are often targeted at tumors. In addition, the term "antibody therapy" according to the invention preferably does not include therapy with antibodies, fragments or derivatives thereof that are conjugated to a therapeutic group, but simply refers to therapy with antibodies, fragments or derivatives thereof, acting by recruiting the patient's immune system to kill tumor cells.

The term "antigen" refers to an agent containing an epitope against which an immune response is to be generated and/or an immune response is directed. The term "antigen" includes, in particular, proteins, peptides, polysaccharides, nucleic acids, especially RNA and DNA, and nucleotides. The term "antigen" also includes agents that become antigenic (and sensitizing) only through transformation (eg, intermediate in the molecule or completion by the body of the protein). The antigen or its processing product is preferably recognized by a T or B cell receptor or an immunoglobulin molecule such as an antibody. In a preferred embodiment, the antigen is a disease-associated antigen, such as a tumor antigen such as CLDN18.2.

In the context of the present invention, the term "tumor antigen" or "tumor-associated antigen" refers to an antigen that is present in tumor cells. Preferably, the antigen is present in the tumor cells, for example, on the surface of the tumor cells. Preferably, the "tumor antigen" is expressed by the tumor cells. In one embodiment, the term "tumor antigen" refers to proteins that are aberrantly expressed in tumor cells compared to normal, ie, non-tumor cells. For example, expression may only be found in tumor cells and not in normal, ie, non-tumor cells, or the level of expression may be higher in tumor cells compared to normal, ie, non-tumor cells. In one embodiment, the term "tumor antigen" refers to proteins that are under normal conditions, specifically expressed in a limited number of tissues and/or organs, or at specific developmental stages, and expressed or aberrantly expressed in one or more tumor or malignant tumor tissues. In the context of the present invention, the tumor antigen is preferably associated with the cell surface of the cancer cell and is preferably not only, or rarely, or at a lower level, expressed in normal tissues and cells. Preferably, according to the invention, the tumor antigen is not expressed in the cell if the expression level is below the detection limit and/or if the expression level is too low to allow binding of antibodies specific for tumor antigens added to the cells. A particularly preferred tumor antigen according to the invention is CLDN18.2.

According to the invention, the term "tumor antigen positive cancer" or "tumor antigen positive tumor" or similar terms means a cancer or tumor comprising cancer or tumor cells expressing tumor antigen, preferably on the surface of said cancer cells or tumor cells. cells. A tumor antigen is expressed on the surface of cells if it is located on the surface of said cells and is available for binding to tumor antigen-specific antibodies added to the cells.

In one preferred embodiment of the invention, the "tumor antigen positive cancer" or "tumor antigen positive tumor" is "CLDN18.2 positive cancer" or "CLDN18.2 positive tumor". According to the invention, the term "CLDN18.2-positive malignancy" or "CLDN18.2-positive tumor" means a malignant neoplasm or tumor, including malignant tumor or tumor cells expressing CLDN18.2, preferably on the surface of said malignant tumor cells or tumor cells.

"Cell surface" is used according to its normal meaning in the art and thus includes the outer surface of a cell that is available for binding by proteins and other molecules.

The term "extracellular portion" in the context of the present invention refers to the portion of a molecule, such as a protein, that faces the extracellular space of a cell and is preferably accessible from outside said cell, for example, to antigen-binding molecules such as antibodies located outside the cell. Preferably, the term refers to one or more extracellular loops or domains, or a fragment thereof.

According to the invention, CLDN18.2 is not significantly expressed in a cell if the expression level is lower compared to expression in gastric cells or gastric tissue. Preferably, the expression level is less than 10%, preferably less than 5%, 3%, 2%, 1%, 0.5%, 0.1% or 0.05% of expression in gastric cells or gastric tissues, or even lower. Preferably, CLDN18.2 is not significantly expressed in a cell if the expression level exceeds the expression level in non-malignant tissue other than the stomach by no more than 2-fold, preferably 1.5-fold, and preferably does not exceed the expression level in said non-malignant tissue. Preferably, CLDN18.2 is not substantially expressed in the cell if the expression level is below the detection limit and/or if the expression level is too low to allow binding by CLDN18.2-specific antibodies added to the cells.

According to the invention, CLDN18.2 is expressed in a cell if the expression level exceeds the expression level in non-cancerous tissue other than the stomach, preferably more than 2 times, preferably 10 times, 100 times, 1000 or 10000 times. Preferably, CLDN18.2 is expressed in a cell if the expression level is above the detection limit and/or if the expression level is high enough to allow binding to CLDN18.2-specific antibodies added to the cells. Preferably, CLDN18.2 expressed in a cell is expressed or displayed on the surface of said cell.

The term "epitope" refers to an antigenic determinant in a molecule, i. to the part of the molecule that is recognized by the immune system, for example, that is recognized by an antibody. For example, epitopes are discrete three-dimensional regions on an antigen that are recognized by the immune system. Epitopes usually consist of reactive surface groups of molecules, such as amino acids or sugar side chains, and usually have specific three-dimensional structural characteristics as well as specific charge characteristics. Conformational and non-conformational epitopes differ in that binding to the former, but not to the latter, is lost in the presence of denaturing solvents. A protein epitope preferably contains a continuous or discontinuous portion of said protein and is preferably 5 to 100, preferably 5 to 50, more preferably 8 to 30, most preferably 10 to 25 amino acids in length, e.g., the epitope may preferably be 8, 9 , 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length.

The term "antibody" includes a glycoprotein containing at least two heavy (H) chains and two light chains (L) linked by disulfide bonds, and any molecule containing the antigen-binding portion of such a glycoprotein. The term "antibody" includes monoclonal antibodies, recombinant antibodies, human antibodies, humanized antibodies, chimeric antibodies, antibody fragments or derivatives, including, but not limited to, single chain antibodies such as scFv and antigen-binding antibody fragments such as Fab and Fab' fragments, and also includes all recombinant forms of antibodies, such as antibodies expressed in prokaryotes, non-glycosylated antibodies, and any antigen-binding antibody fragments and derivatives described herein. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) interspersed with regions that are more conserved and called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from amino to carboxy in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. Antibody constant regions can mediate immunoglobulin binding to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.

As used herein, the term "monoclonal antibody" refers to a preparation of antibody molecules with a single molecular composition. A monoclonal antibody exhibits a single binding specificity and affinity. In one embodiment, the monoclonal antibodies are generated by a hybridoma that includes a B cell derived from a non-human animal, such as a mouse, fused to an immortalized cell.

As used herein, the term "recombinant antibody" includes all antibodies that have been produced, expressed, generated, or isolated by recombinant means, such as (a) antibodies isolated from an animal (e.g., mouse) that is transgenic or transchromosomal for immunoglobulin genes , or a hybridoma derived therefrom, (b) antibodies isolated from a host cell transformed to express the antibody, e.g. from a transfectoma, (c) antibodies isolated from a library of recombinant combinatorial antibodies, and (d) antibodies obtained, expressed, created or isolated by any other means that involve splicing immunoglobulin gene sequences with other DNA sequences.

As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from germline human immunoglobulin sequences. Human antibodies may include amino acid residues not encoded by germline human immunoglobulin sequences (eg, mutations introduced by in vitro random or site-specific mutagenesis or in vivo somatic mutation).

The term "humanized antibody" refers to a molecule having an antigen binding site that is essentially derived from an immunoglobulin of a non-human species, in which the remaining structure of the immunoglobulin molecule is based on the structure and/or sequence of a human immunoglobulin. The antigen binding site may either contain complete variable domains fused to constant domains, or only complementarity determining regions (CDRs) grafted into the appropriate framework regions in the variable domains. Antigen binding sites may be wild-type or modified with one or more amino acid substitutions, eg modified to more closely resemble human immunoglobulins. Some forms of humanized antibodies retain all CDR sequences (eg, a humanized mouse antibody that contains all six CDRs from a mouse antibody). Other forms have one or more CDRs that vary from the parent antibody.

The term "chimeric antibody" refers to those antibodies where one portion of each of the heavy and light chain amino acid sequences is homologous to corresponding sequences in antibodies derived from the same species or belonging to a particular class, while the remaining chain segment is homologous to corresponding sequences in another species or class. . Typically, the variable region of both the light and heavy chains mimics the variable regions of antibodies derived from one mammalian species, while the constant portions are homologous to antibody sequences derived from another organism. One distinct advantage of such chimeric forms is that the variable region can be conveniently derived from currently known sources using readily available B cells or hybridomas from non-human hosts in combination with constant regions derived from e.g. human cells. Although the variable region has the advantage of ease of preparation and the specificity is not affected by the source, their human constant region is less likely to elicit an immune response in a human when administered with an antibody than a constant region from a non-human source. However, this definition is not limited to this particular example.

Antibodies can be obtained from a variety of species, including, but not limited to, mice, rats, rabbits, guinea pigs, and humans.

Antibodies described herein include IgA such as IgA1 or IgA2, IgG1, IgG2, IgG3, IgG4, IgE, IgM and IgD antibodies. In various embodiments of the invention, the antibody is an IgG1 antibody, more specifically an IgG1, kappa or IgG1, lambda isotype (i.e., IgG1, κ, λ), an IgG2a antibody (e.g., IgG2a, κ, λ), an IgG2b antibody (e.g., IgG2b, κ, λ), IgG3, antibody (eg IgG 3, κ, λ) or IgG4 antibody (eg IgG4, κ, λ).

As used herein, the term "heterologous antibody" is defined in relation to a transgenic organism that produces such an antibody. This term refers to an antibody having an amino acid sequence or a nucleic acid coding sequence corresponding to that found in an organism that does not match the transgenic organism and is usually derived from a species other than the transgenic organism.

As used herein, the term "heterohybrid antibody" refers to an antibody having light and heavy chains from different organisms. For example, an antibody having a human heavy chain linked to a mouse light chain is a heterohybrid antibody.

The antibodies described herein are preferably isolated. "Isolated antibody", as used herein, is intended to mean an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to a tumor antigen is substantially free of antibodies that specifically bind antigens of another tumor antigen). An isolated antibody that specifically binds to an epitope, isoform, or variant of a human tumor antigen may, however, be cross-reactive with other related antigens, eg from other species (eg, tumor antigen homologues). In addition, the isolated antibody may be substantially free of other cellular material and/or chemicals. In one embodiment of the invention, a combination of "isolated" monoclonal antibodies refers to antibodies having different specificities and combined in a well-defined composition or mixture.

The terms "antigen-binding portion" of an antibody (or simply "binding portion") or "antigen-binding fragment" of an antibody (or simply "binding fragment") or similar terms refer to one or more antibody fragments that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full length antibody. Examples of binding fragments included in the term "antigen-binding portion" of an antibody include (i) Fab fragments, monovalent fragments consisting of VL, VH, CL and CH domains; (ii) F(ab') ₂ fragments, divalent fragments containing two Fab fragments connected by a disulfide bridge in the hinge region; (iii) Fd fragments consisting of VH and CH domains; (iv) Fv fragments consisting of the VL and VH domains of one antibody arm, (v) dAb fragments (Ward et al., (1989) Nature 341: 544-546) which consist of a VH domain; (vi) dedicated complementarity determination regions (CDRs); and (vii) combinations of two or more dedicated CDRs, which may optionally be joined by a synthetic linker. In addition, although the two fragment regions Fv, VL and VH are encoded by separate genes, they can be combined, using recombinant methods, with a synthetic linker, which allows to obtain a single protein chain in which the VL and VH pairs form monovalent molecules (known as single-stranded Fv ( scFv); Such single chain antibodies are also intended to be included in the term "antigen binding fragment" of an antibody. Another example are immunoglobulin-binding domain fusion proteins comprising (i) a binding domain polypeptide that is fused to an immunoglobulin hinge polypeptide, (ii) an immunoglobulin heavy chain CH2 constant region fused to the hinge, and (iii) a heavy chain CH3 constant region. immunoglobulin fused to the CH2 constant region. The binding domain polypeptide can be a heavy chain variable region or a light chain variable region. Binding domain and immunoglobulin fusion proteins are further described in US 2003/0118592 and US 2003/0133939. These antibody fragments are prepared by conventional methods known to those skilled in the art and the fragments are screened for utility in the same manner as intact antibodies.

The term "binding domain" characterizes in connection with the present invention a structure, such as an antibody, that binds/interacts with a given target structure/antigen/epitope. Thus, the binding domain in accordance with the invention means "antigen interaction site".

All antibodies and antibody derivatives, such as antibody fragments, as described herein for the purposes of the invention, are encompassed by the term "antibody". The term "antibody derivatives" refers to any modified form of an antibody, such as a conjugate of an antibody and another agent or antibody, or an antibody fragment.

Naturally occurring antibodies tend to be monospecific, meaning they bind to a single antigen. The present invention includes antibodies that bind to a target cell (by interacting with a tumor antigen) and a second entity, such as a cytotoxic cell (eg, by interacting with the CD3 receptor). The antibodies of the present invention may be bispecific or multispecific, such as trispecific, tetraspecific, and so on.

The term "bispecific molecule" is intended to include an agent that has two different binding specificities. For example, the molecule may bind to (a) a cell surface antigen or (b) a receptor such as an Fc receptor on the surface of an effector cell. The term "multispecific molecule" is intended to include an agent that has more than two different binding specificities. For example, the molecule can bind to or interact with (a) a cellular antigen, (b) a receptor, such as an Fc receptor on the surface of an effector cell, and (c) at least one other component. Accordingly, the term "tumor antigen antibody" includes, but is not limited to, bispecific, trispecific, tetraspecific, and other multispecific molecules that target the tumor antigen and other targets such as Fc receptors on effector cells. The term "bispecific antibodies" also includes diabodies. Diabodies are bivalent, bispecific antibodies in which the VH and VL domains are expressed on the same polypeptide chain, but using a linker that is too short to allow pairing between two domains on the same chain, thereby causing the domains to pair with complementary other strand domains and create two antigen binding sites (see, e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448; Poljak, R. J., et al. (1994 ) Structure 2: 1121-1123).

In accordance with the invention, an antibody may exert a therapeutic effect by recruiting the patient's immune system to kill tumor cells and/or via a therapeutic moiety or agent linked to the antibody. For the purposes of the present invention, such antibody conjugates may be considered to be encompassed by the term "chemotherapeutic agent", while antibodies that provide a therapeutic effect by recruiting the patient's immune system to kill tumor cells cannot be so considered.

In the context of the present invention, the antibody is preferably capable of acting by recruiting the patient's immune system to kill tumor cells, i.e. the antibody, in particular when bound to its target, such as a tumor antigen on the affected cell, produces the immune effect described herein. Preferably, said immune effector functions are directed against cells, such as cancer cells, bearing a tumor antigen, such as CLDN18.2, on their surface.

The term "immune effector functions" in the context of the present invention includes any functions mediated by components of the immune system that result, for example, in inhibition of tumor growth and/or inhibition of tumor development, including inhibition of tumor spread and metastasis. Preferably, the functions of the immune effector lead to the destruction of cancer cells. Such functions include complementary-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), induction of apoptosis in tumor antigen-bearing cells, cytolysis of tumor antigen-bearing cells, and/or inhibition of cell proliferation. carrying the tumor antigen. Binding agents may also act simply by binding to tumor antigens on the surface of the cancer cell. For example, antibodies can block the function of a tumor antigen or induce apoptosis only by binding to a tumor antigen on the surface of a cancer cell.

Antibody-dependent cell-mediated cytotoxicity

ADCC describes the ability of effector cells, in particular lymphocytes, which preferentially require the target cell to be labeled with an antibody, to kill cells.

ADCC preferably occurs when antibodies bind to antigens on tumor cells and antibody Fc domains interact with Fc receptors (FcRs) on the surface of immune effector cells. Several families of Fc receptors have been identified, and specific cell populations characteristically express certain Fc receptors. ADCC can be seen as a mechanism to directly induce a variable degree of immediate tumor destruction, leading to antigen presentation and induction of tumor-induced T cell responses. Preferably, in vivo induction of ADCC will result in tumor-induced T cell and host antibody responses.

Complement dependent cytotoxicity

CDC is another way of killing cells that can be driven by antibodies. IgM is the most efficient complement activation isotype. IgG1 and IgG3 are also very effective in driving CDC through the classical complement pathway. Preferably, in this cascade, the formation of antigen-antibody complexes results in the exposure of multiple C1q binding sites in close proximity to the _CH 2 domains of participating antibody molecules, such as IgG molecules (C1q is one of the three subcomponents of C1 complement). Preferably, these exposed C1q binding sites convert a previously low affinity C1q-IgG interaction into a high avidity interaction that triggers a cascade of events involving a number of other complement proteins and results in proteolytic release of C3a and C5a effector cell chemotactic/activating agents. Preferably, the complement cascade ends with the formation of a membrane attack complex, which creates pores in the cell membrane to facilitate the free passage of water and solutes into and out of the cell.

In order to inhibit tumor growth and/or tumor development, according to the invention, an antibody may be conjugated to a therapeutic moiety or agent, such as a cytotoxin, a drug (eg, an immunosuppressant), or a radioisotope. A cytotoxin or cytotoxic agent includes any agent that is detrimental to a cell and specifically kills cells. Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracindione, mitoxantrone, mithramycin, actinomycin D, amanitin, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine , lidocaine, propranolol and puromycin, as well as their analogues or homologues. Suitable therapeutic agents for the formation of antibody conjugates include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine ( BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cisplatin (ii) cisplatin (DDP)), anthracyclines (eg, daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (eg, dactinomycin (formerly actinomycin ), bleomycin, mithramycin, and anthramycin (AMC), and antimitotic agents (e.g., vincristine and vinblastine). In a preferred embodiment, the therapeutic agent is a cytotoxic agent or a radiotoxic agent. In another embodiment, the therapeutic agent is an immunosuppressant. In yet another embodiment, the therapeutic agent is is GM-CSF In a preferred embodiment, the therapeutic agent is is doxorubicin, cisplatin, bleomycin, sulfate, carmustine, chlorambucil, cyclophosphamide, or ricin A.

Antibodies can also be conjugated to a radioisotope, such as iodine-131, yttrium-90, or indium-111, to produce cytotoxic radiopharmaceuticals.

The antibody conjugates of the invention can be used to modify this biological response and the drug portion should not be construed as being limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide having the biological activity of interest. Such proteins may include, for example, an enzymatically active toxin or active fragment thereof, such as abrin, ricin A, Pseudomonas aeruginosa exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor or interferon γ; or, biological response modifiers such as, for example, lymphokines, interleukin-1 ("IL-1"), interleukin-2 ("IL-2"), interleukin-6 ("IL-6"), granulocyte-macrophage colony-stimulating factor ("GM-CSF"), granulocyte colony stimulating factor ("G-CSF") or other growth factors.

Methods for conjugating such a therapeutic moiety to antibodies are well known, see, for example, Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery", in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pincheraet al. (eds.), pp. 475-506 (1985); "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev. 62: 119-58 (1982).

The term "anti-tumor antigen antibody" or similar terms refers to an antibody directed to or capable of binding to a tumor antigen. The term "binding" according to the invention preferably refers to specific binding.

In accordance with the present invention, an antibody is capable of binding to a predetermined target if it has significant affinity for said predetermined target and binds to said predetermined target in standard assays. "Affinity" or "binding affinity" is often measured by the equilibrium dissociation constant (K _D ). Preferably, the term "significant affinity" refers to binding to a given target with a dissociation constant (K _D ) of 10 ^-5 M or less, 10 ^-6 M or less, 10 ^-7 M or less, 10 ^-8 M or less, 10 ^-9 M or less, 10 ^-10 M or less, 10 ^-11 M or less, or 10 ^-12 M or less.

An antibody is not (substantially) capable of binding to a target if it does not have significant affinity for said target and does not bind significantly, in particular, does not bind in a detectable manner to said target in standard assays. Preferably, the antibody does not bind to said target if it is present at a concentration of up to 2, preferably 10, more preferably 20, in particular 50 or 100 μg/ml or higher. Preferably, if the antibody has no significant affinity for the target, if it binds to said target with a K _D that is at least 10 times, 100 times, 10 ³ times, 10 ⁴ times, 10 ⁵ times, or 10 ⁶ times higher than the K _D for binding to a predetermined target to which the antibody is able to bind. For example, if the K _D for binding an antibody to a target for which the antibody is able to bind is 10 ^-7 M, the K _D for binding to a target for which the antibody has no significant affinity would be at least 10 ^-6 M, 10 ^-5 M, 10 ^-4 M, 10 ^-3 M, 10 ^-2 M or 10 ^-1 M.

An antibody is specific for a given target if it is able to bind to said predefined target while it is not able to bind to other targets, i.e. has no significant affinity for other targets and cannot bind significantly to other targets in standard assays. According to the invention, an antibody is specific for a tumor antigen if it is able to bind to the tumor antigen but is not (substantially) able to bind to other targets. Preferably, the antibody is specific for the tumor antigen if the binding affinity for and binding to such other targets is not significantly greater than the affinity for or binding to proteins not bound to the tumor antigen, such as bovine serum albumin (BSA), casein, human serum albumin (HSA) or transmembrane proteins not associated with the tumor antigen, such as MHC molecules or the transferrin receptor or any other specified polypeptide. Preferably, an antibody is specific for a predetermined target if it binds to said entity with a K _D that is at least 10 times, 100 times, 10 ³ times, 10 ⁴ times, 10 ⁵ times, or 10 ⁶ times less than than K _D binding to a target for which it is not specific. For example, if the K _D for binding of an antibody to a target for which it is specific is ^10-7 M, the K _D for binding to a target for which it is not specific will be at least 10 ^-6 M, 10 ^{- 5} M, 10 ^-4 M, 10 ^-3 M, 10 ^-2 M or 10 ^-1 M.

Binding of an antibody to a target can be determined experimentally using any suitable method; see, for example, Berzofsky et al., "Antibody-Antigen Interactions" In Fundamental Immunology, Paul, WE, Ed., Raven Press New York, NY (1984), Kuby, Janis Immunology, WH Freeman and Company New York, NY (1992) and the methods described herein. Affinities can be easily determined using conventional methods such as equilibrium dialysis; using the BIAcore 2000 instrument, using the general procedures specified by the manufacturer; by radioimmunoassay using a radioactively labeled antigen; or in other manner known to the person skilled in the art. Affinity data can be analyzed, for example, by the method of Scatchard et al., Ann NY Acad. ScL, 51:660 (1949). The measured affinity of a particular antibody-antigen interaction may vary when measured under different conditions, eg salt concentration, pH. Thus, measurements of affinity and other antigen-binding parameters, eg K _D , IC ₅₀ , are preferably made with standardized antibody and antigen solutions and a standardized buffer.

As used herein, the term "isotype" refers to the class of antibodies (eg, IgM or IgG1) that is encoded by heavy chain constant region genes.

As used herein, the term "isotype switching" refers to the phenomenon by which the class or isotype of an antibody changes from one Ig class to one of the other Ig classes.

The term "naturally occurring" as used herein to apply to an object refers to the fact that the object can be found in nature. For example, a polypeptide or polynucleotide sequence present in an organism (including viruses) that can be isolated from a source in nature and that has not been intentionally modified by humans in a laboratory is naturally occurring.

As used herein, the term "rearranged" refers to a heavy chain or light chain immunoglobulin locus configuration where the V segment is located immediately adjacent to the D-J or J segment in a conformation encoding a substantially complete VH or VL domain, respectively. The rearranged immunoglobulin (antibody) gene locus can be identified by comparison with germline DNA; the rearranged locus will have at least one recombined heptamer/nonamer homology element.

The term "unrearranged" or "germline configuration", as used herein in relation to a V segment, refers to a configuration in which the V segment is not recombined to be directly adjacent to the D or J segment.

Preferably, binding of an antibody against a tumor antigen to cells expressing the tumor antigen induces or mediates the killing of cells expressing the tumor antigen. Cells expressing the tumor antigen are preferably cancer cells, and are in particular cancer cells described herein. Preferably, the antibody induces or mediates cell killing by inducing one or more of complement-dependent cytotoxicity (CDC)-mediated lysis, antibody-dependent cellular cytotoxicity (ADCC)-mediated lysis, apoptosis, and inhibition of proliferation of tumor antigen-expressing cells. Preferably, ADCC-mediated cell lysis occurs in the presence of effector cells, which in specific embodiments are selected from the group consisting of monocytes, mononuclear cells, NK cells and PMNs. Inhibition of cell proliferation can be measured in vitro by determining cell proliferation in an assay using bromodeoxyuridine (5-bromo-2'-deoxyuridine, BrdU). BrdU is a synthetic nucleoside that is analogous to thymidine and can be incorporated into the newly synthesized DNA of replicating cells (during the S-phase of the cell cycle), replacing thymidine during DNA replication. Detection of an incorporated chemical using, for example, antibodies specific for BrdU indicates cells that are actively replicating their DNA.

In preferred embodiments, the antibodies described herein may be characterized by one or more of the following properties:

a) tumor antigen specificity;

b) a binding affinity for the tumor antigen of about 100 nM or less, preferably about 5-10 nM or less, and more preferably about 1-3 nM or less,

c) the ability to induce or mediate CDC on tumor antigen positive cells;

d) the ability to induce or mediate ADCC on tumor antigen positive cells;

e) the ability to inhibit the growth of tumor antigen positive cells;

f) the ability to induce apoptosis of tumor antigen positive cells.

In one embodiment, an antibody against a tumor antigen has the ability to bind to an epitope present on the tumor antigen, preferably an epitope located within the extracellular domains of the tumor antigen. Preferably, the anti-tumor antigen antibody is specific for the tumor antigen. Preferably, the anti-tumor antigen antibody binds to a tumor antigen expressed on the cell surface. In particularly preferred embodiments, the anti-tumor antigen antibody binds to native tumor antigen epitopes present on the surface of living cells.

According to the invention, an antibody capable of binding to CLDN18.2 or an antibody against CLDN18.2 is an antibody capable of binding to an epitope present in CLDN18.2, preferably an epitope located in the extracellular domains of CLDN18.2, in particular the first extracellular domain , preferably at amino acid positions 29-78 in CLDN18.2. In specific embodiments, an antibody capable of binding to CLDN18.2 is an antibody that is capable of binding to (i) an epitope on CLDN18.2 that is not present on CLDN18.1, preferably SEQ ID NOS: 3, 4 and 5, (ii) an epitope located in CLDN18.2 loop 1, preferably SEQ ID NO: 8, (iii) an epitope located in CLDN18.2 loop 2, preferably SEQ ID NO: 10, (iv) an epitope located in CLDN18.2- loop D3, preferably SEQ ID NO: 11, (v) an epitope that spans the CLDN18.2 loop 1 and CLDN18.2 loop D3, or (vi) a non-glycosylated epitope located on the CLDN18.2 loop D3, preferably SEQ ID NO:9.

According to the invention, an antibody having the ability to bind to CLDN18.2 is preferably an antibody having the ability to bind to CLDN18.2 but not to CLDN18.1. Preferably, the antibody having the ability to bind to CLDN18.2 is specific for CLDN18.2. Preferably, the antibody having the ability to bind to CLDN18.2 is an antibody having the ability to bind to CLDN18.2 expressed on the cell surface. In preferred embodiments, an antibody capable of binding to CLDN18.2 binds to native CLDN18.2 epitopes present on the surface of living cells. Preferably, an antibody with the ability to bind to CLDN18.2 binds to one or more peptides selected from the group consisting of SEQ ID NOs: 1, 3-11, 44, 46 and 48-50. Preferably, the antibody capable of binding to CLDN18.2 is specific for the aforementioned proteins, peptides or immunogenic fragments or derivatives thereof. An antibody having the ability to bind to CLDN18.2 can be obtained by a method comprising the step of immunizing an animal with a protein or peptide containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3-11, 44, 46 and 48-50 , or a nucleic acid or host cell expressing said protein or peptide. Preferably, the antibody binds to cancer cells, in particular to cells of the above types of cancer, and preferably does not substantially bind to cells that are not cancer cells.

Preferably, binding of an antibody capable of binding to CLDN18.2 to cells expressing CLDN18.2 induces or mediates the killing of cells expressing CLDN18.2. Cells expressing CLDN18.2 are preferably cancer cells and are specifically selected from the group consisting of stomach, esophagus, pancreas, lung, ovary, colon, liver, head and neck, and gallbladder cancer cells. Preferably, the antibody induces or mediates cell killing by inducing one or more of complement-dependent cytotoxicity (CDC)-mediated lysis, antibody-dependent cellular cytotoxicity (ADCC)-mediated lysis, apoptosis, and inhibition of proliferation of cells expressing CLDN18.2. Preferably, ADCC-mediated cell lysis occurs in the presence of effector cells, which in specific embodiments are selected from the group consisting of monocytes, mononuclear cells, NK cells and PMNs.

In preferred embodiments, an antibody having the ability to bind to CLDN18.2 may be characterized by one or more of the following properties:

a) specificity for CLDN18.2;

b) a binding affinity for CLDN18.2 of about 100 nM or less, preferably about 5-10 nM or less, and more preferably about 1-3 nM or less,

c) the ability to induce or mediate CDC in CLDN18.2 positive cells;

d) the ability to induce or mediate ADCC in CLDN18.2 positive cells;

e) the ability to inhibit the growth of CLDN18.2 positive cells;

f) the ability to induce apoptosis of CLDN18.2 positive cells.

In a particularly preferred embodiment, an antibody with the ability to bind to CLDN18.2 is obtained using a hybridoma deposited in the DSMZ (Mascheroder Weg 1b, 31824 Braunschweig, Germany; new address: Inhoffenstr. 7B, 31824 Braunschweig, Germany) and has the following designation and account room:

a. 182-D1106-055, DSM account number ACC2737, deposited October 19, 2005

b. 182-D1106-056, DSM account number ACC2738, deposited October 19, 2005

c. 182-D1106-057, DSM account number ACC2739, deposited October 19, 2005

d. 182-D1106-058, DSM account number ACC2740, deposited October 19, 2005

e. 182-D1106-059, DSM account number ACC2741, deposited October 19, 2005

f. 182-D1106-062, DSM account number ACC2742, deposited October 19, 2005,

g. 182-D1106-067, DSM account number ACC2743, deposited October 19, 2005

h. 182-D758-035, DSM account number ACC2745, deposited November 17, 2005.

i. 182-D758-036, DSM account number ACC2746, deposited November 17, 2005.

j. 182-D758-040, DSM account number ACC2747, deposited November 17, 2005.

k. 182-D1106-061, DSM account number ACC2748, deposited November 17, 2005

l. 182-D1106-279, DSM account number ACC2808, deposited October 26, 2006.

m. 182-D1106-294, DSM account number ACC2809, deposited October 26, 2006,

n. 182-D1106-362, DSM account number ACC2810, deposited October 26, 2006.

Preferred antibodies according to the invention are those that are and are derived from the hybridomas described above; those. 37G11 for 182-D1106-055, 37H8 for 182-D1106-056, 38G5 for 182-D1106-057, 38H3 for 182-D1106-058, 39F11 for 182-D1106-059, 43A11 for 182- D1106-062, 61C2 for 182-D1106-067, 26B5 for 182-D758-035, 26D12 for 182-D758-036, 28D10 for 182-D758-040, 42E12 for 182-D1106-061, 125E1 in the case of 182-D1106-279, 163E12 in the case of 182-D1106-294 and 175D10 in the case of 182-D1106-362; and their chimeric and humanized forms.

In one embodiment, the antibody having the ability to bind to CLDN18.2 is an antibody selected from the group consisting of (i) an antibody produced and/or derived from a clone deposited under accession number DSM ACC2737, DSM ACC2738, DSM ACC2739, DSM ACC2740 , DSM ACC2741, DSM ACC2742, DSM ACC2743, DSM ACC2745, DSM ACC2746, DSM ACC2747, DSM ACC2748, DSM ACC2808, DSM ACC2809, or DSM ACC2810, (ii) an antibody that is a chimerized or humanized form of an antibody (i), (iii) an antibody having the specificity of an antibody (i), and (iv) an antibody comprising an antigen-binding portion or an antigen-binding site, in particular a variable region, of the antibody (i) and preferably with the specificity of the antibody (i).

Preferred chimerized antibodies and their sequences are shown in the following table.

clone mAb Isotype Variable region Chimerized antibody heavy chain 43A11 182-D1106-062 IgG2a SEQ ID NO:29 SEQ ID NO:14 163E12 182-D1106-294 IgG3 SEQ ID NO:30 SEQ ID NO:15 125E1 182-D1106-279 IgG2a SEQ ID NO:31 SEQ ID NO:16 166E2 182-D1106-308 IgG3 SEQ ID NO:33 SEQ ID NO:18 175D10 182-D1106-362 IgG1 SEQ ID NO:32 SEQ ID NO:17 45C1 182-D758-187 IgG2a SEQ ID NO:34 SEQ ID NO:19 light chain 43A11 182-D1106-062 IgK SEQ ID NO:36 SEQ ID NO:21 163E12 182-D1106-294 IgK SEQ ID NO:35 SEQ ID NO:20 125E1 182-D1106-279 IgK SEQ ID NO:37 SEQ ID NO:22 166E2 182-D1106-308 IgK SEQ ID NO:40 SEQ ID NO:25 175D10 182-D1106-362 IgK SEQ ID NO:39 SEQ ID NO:24 45C1 182-D758-187 IgK SEQ ID NO:38 SEQ ID NO:23 45C1 182-D758-187 IgK SEQ ID NO:41 SEQ ID NO:26 45C1 182-D758-187 IgK SEQ ID NO:42 SEQ ID NO:27 45C1 182-D758-187 IgK SEQ ID NO:43 SEQ ID NO:28

In preferred embodiments, antibodies, in particular chimerized forms of antibodies according to the invention, include antibodies comprising a heavy chain constant region (CH) containing an amino acid sequence derived from a human heavy chain constant region, such as the amino acid sequence shown in SEQ ID NO : 13 or a fragment of it. In other preferred embodiments, antibodies, in particular chimerized forms of antibodies in accordance with the invention, include antibodies containing a light chain constant region (CL) containing an amino acid sequence derived from a human light chain constant region, such as the amino acid sequence shown in SEQ ID NO : 12 or its fragment. In a specific preferred embodiment, antibodies, in particular chimerized forms of antibodies according to the invention, include antibodies that contain a CH containing an amino acid sequence derived from human CH, such as the amino acid sequence shown in SEQ ID NO: 13 of it or a fragment, and which include a CL containing an amino acid sequence derived from human CL, such as the amino acid sequence shown in SEQ ID NO: 12, or a fragment thereof.

In one embodiment, the antibody capable of binding to CLDN18.2 is a chimeric mouse/human IgG1 monoclonal antibody comprising a mouse kappa light chain variable region, a human kappa light chain constant region of the Km(3) allotype, a mouse heavy chain variable region, a constant human IgG1 region, allotype G1m (3).

In some preferred embodiments, chimerized forms of antibodies include antibodies containing a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 14, 15, 16, 17, 18, 19, 51 and a fragment thereof and/or containing a light chain , comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 20, 21, 22, 23, 24, 25, 26, 27, 28 and a fragment thereof.

In some preferred embodiments, chimerized forms of antibodies include antibodies containing a combination of heavy chains and light chains selected from the following options (i)-(ix):

(i) the heavy chain comprises the amino acid sequence shown in SEQ ID NO: 14 or a fragment thereof, and the light chain contains the amino acid sequence shown in SEQ ID NO: 21 or a fragment thereof,

(ii) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 15 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 20 or a fragment thereof,

(iii) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 16 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 22 or a fragment thereof,

(iv) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 18 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 25 or a fragment thereof,

(v) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 17 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 24 or a fragment thereof,

(vi) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 19 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 23 or a fragment thereof,

(vii) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 19 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 26 or a fragment thereof,

(viii) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 19 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 27 or a fragment thereof,

(ix) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 19 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 28 or a fragment thereof, and

(x) the heavy chain contains the amino acid sequence represented by SEQ ID NO: 51 or a fragment thereof, and the light chain contains the amino acid sequence represented by SEQ ID NO: 24 or a fragment thereof.

Especially preferred is the antibody according to (v) or (x).

"Fragment" or "fragment of an amino acid sequence" as used above refers to a portion of an antibody sequence, i.e., a sequence that represents an antibody sequence truncated at the N- and/or C-terminus, which, when the sequence of said antibody is replaced in the antibody, retains binding said antibody with CLDN18.2 and preferably the function of said antibody as described herein, eg, CDC-mediated lysis or ADCC-mediated lysis. Preferably, the amino acid sequence fragment contains at least 80%, preferably at least 90%, 95%, 96%, 97%, 98% or 99% of the amino acid residues from said amino acid sequence. An amino acid sequence fragment selected from the group consisting of SEQ ID NOs: 14, 15, 16, 17, 18, 19, 51, 20, 21, 22, 23, 24, 25, 26, 27 and 28, preferably refers to the specified sequences where 17, 18, 19, 20, 21, 22, or 23 amino acids at the N-terminus are deleted.

In a preferred embodiment, an antibody capable of binding to CLDN18.2 comprises a heavy chain variable region (VH) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 29, 30, 31, 32, 33, 34 and fragments thereof .

In a preferred embodiment, an antibody capable of binding to CLDN18.2 comprises a light chain variable region (VL) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 35, 36, 37, 38, 39, 40, 41, 42, 43 and their fragments.

In some preferred embodiments, an antibody capable of binding to CLDN18.2 comprises a combination of a heavy chain variable region (VH) and a light chain variable region (VL) selected from the following options (i)-(ix):

(i) VH contains the amino acid sequence represented by SEQ ID NO: 29 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 36 or a fragment thereof,

(ii) VH contains the amino acid sequence represented by SEQ ID NO: 30 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 35 or a fragment thereof,

(iii) VH contains the amino acid sequence represented by SEQ ID NO: 31 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 37 or a fragment thereof,

(iv) VH contains the amino acid sequence represented by SEQ ID NO: 33 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 40 or a fragment thereof,

(v) VH contains the amino acid sequence represented by SEQ ID NO: 32 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 39 or a fragment thereof,

(vi) VH contains the amino acid sequence represented by SEQ ID NO: 34 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 38 or a fragment thereof,

(vii) VH contains the amino acid sequence represented by SEQ ID NO: 34 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 41 or a fragment thereof,

(viii) VH contains the amino acid sequence represented by SEQ ID NO: 34 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 42 or a fragment thereof,

(ix) VH contains the amino acid sequence represented by SEQ ID NO: 34 or a fragment thereof, and VL contains the amino acid sequence represented by SEQ ID NO: 43 or a fragment thereof.

Especially preferred is the antibody according to (v).

According to the invention, the term "fragment" refers in particular to one or more complementarity determining regions (CDRs), preferably at least the CDR3 variable region of the heavy chain variable region (VH) and/or the light chain variable region ( VL). In one embodiment, said one or more complementarity determining regions (CDRs) are selected from a set of complementarity determining regions CDR1, CDR2, and CDR3. In a particularly preferred embodiment, the term "fragment" refers to the complementarity-determining regions of the heavy chain variable region (VH) and/or light chain variable region (VL) CDR1, CDR2 and CDR3.

In a preferred embodiment, the antibody capable of binding to CLDN18.2 contains a VH containing a set of CDR1, CDR2, and CDR3 complementarity-determining regions selected from the following options (i)-(vi):

(i) CDR1: positions 45-52 of SEQ ID NO: 14, CDR2: positions 70-77 of SEQ ID NO: 14, CDR3: positions 116-125 of SEQ ID NO: 14,

(ii) CDR1: positions 45-52 of SEQ ID NO: 15, CDR2: positions 70-77 of SEQ ID NO: 15, CDR3: positions 116-126 of SEQ ID NO: 15,

(iii) CDR1: positions 45-52 of SEQ ID NO: 16, CDR2: positions 70-77 of SEQ ID NO: 16, CDR3: positions 116-124 of SEQ ID NO: 16,

(iv) CDR1: positions 45-52 of SEQ ID NO: 17, CDR2: positions 70-77 of SEQ ID NO: 17, CDR3: positions 116-126 of SEQ ID NO: 17,

(v) CDR1: positions 44-51 of SEQ ID NO: 18, CDR2: positions 69-76 of SEQ ID NO: 18, CDR3: positions 115-125 of SEQ ID NO: 18 and

(vi) CDR1: positions 45-53 of SEQ ID NO: 19, CDR2: positions 71-78 of SEQ ID NO: 19, CDR3: positions 117-128 of SEQ ID NO: 19.

In a preferred embodiment, an antibody having the ability to bind to CLDN18.2 comprises a VL containing a set of CDR1, CDR2 and CDR3 complementarity determining regions selected from the following embodiments (i)-(ix):

(i) CDR1: positions 47-58 of SEQ ID NO: 20, CDR2: positions 76-78 of SEQ ID NO: 20, CDR3: positions 115-123 of SEQ ID NO: 20,

(ii) CDR1: positions 49-53 of SEQ ID NO: 21, CDR2: positions 71-73 of SEQ ID NO: 21, CDR3: positions 110-118 of SEQ ID NO: 21,

(iii) CDR1: positions 47-52 of SEQ ID NO: 22, CDR2: positions 70-72 of SEQ ID NO: 22, CDR3: positions 109-117 of SEQ ID NO: 22,

(iv) CDR1: positions 47-58 of SEQ ID NO: 23, CDR2: positions 76-78 of SEQ ID NO: 23, CDR3: positions 115-123 of SEQ ID NO: 23,

(v) CDR1: positions 47-58 of SEQ ID NO: 24, CDR2: positions 76-78 of SEQ ID NO: 24, CDR3: positions 115-123 of SEQ ID NO: 24,

(vi) CDR1: positions 47-58 of SEQ ID NO: 25, CDR2: positions 76-78 of SEQ ID NO: 25, CDR3: positions 115-122 of SEQ ID NO: 25,

(vii) CDR1: positions 47-58 of SEQ ID NO: 26, CDR2: positions 76-78 of SEQ ID NO: 26, CDR3: positions 115-123 of SEQ ID NO: 26,

(viii) CDR1: positions 47-58 of SEQ ID NO: 27, CDR2: positions 76-78 of SEQ ID NO: 27, CDR3: positions 115-123 of SEQ ID NO: 27 and

(ix) CDR1: positions 47-52 of SEQ ID NO: 28, CDR2: positions 70-72 of SEQ ID NO: 28, CDR3: positions 109-117 of SEQ ID NO: 28.

In a preferred embodiment, an antibody capable of binding to CLDN18.2 comprises a combination of VH and VL, each containing a set of CDR1, CDR2, and CDR3 complementarity-determining regions selected from the following embodiments (i)-(ix):

(i) VH: CDR1: positions 45-52 SEQ ID NO: 14, CDR2: positions 70-77 SEQ ID NO: 14, CDR3: positions 116-125 SEQ ID NO: 14, VL: CDR1: positions 49 -53 SEQ ID NO: 21, CDR2: positions 71-73 SEQ ID NO: 21, CDR3: positions 110-118 SEQ ID NO: 21,

(ii) VH: CDR1: positions 45-52 SEQ ID NO: 15, CDR2: positions 70-77 SEQ ID NO: 15, CDR3: positions 116-126 SEQ ID NO: 15, VL: CDR1: positions 47 -58 SEQ ID NO: 20, CDR2: positions 76-78 SEQ ID NO: 20, CDR3: positions 115-123 SEQ ID NO: 20,

(iii) VH: CDR1: positions 45-52 SEQ ID NO: 16, CDR2: positions 70-77 SEQ ID NO: 16, CDR3: positions 116-124 SEQ ID NO: 16, VL: CDR1: positions 47 -52 SEQ ID NO: 22, CDR2: positions 70-72 SEQ ID NO: 22, CDR3: positions 109-117 SEQ ID NO: 22,

(iv) VH: CDR1: positions 44-51 SEQ ID NO: 18, CDR2: positions 69-76 SEQ ID NO: 18, CDR3: positions 115-125 SEQ ID NO: 18, VL: CDR1: positions 47-58 SEQ ID NO: 25, CDR2: positions 76-78 SEQ ID NO: 25, CDR3: positions 115-122 SEQ ID NO: 25,

(v) VH: CDR1: positions 45-52 SEQ ID NO: 17, CDR2: positions 70-77 SEQ ID NO: 17, CDR3: positions 116-126 SEQ ID NO: 17, VL: CDR1: positions 47 -58 SEQ ID NO: 24, CDR2: positions 76-78 SEQ ID NO: 24, CDR3: positions 115-123 SEQ ID NO: 24,

(vi) VH: CDR1: positions 45-53 SEQ ID NO: 19, CDR2: positions 71-78 SEQ ID NO: 19, CDR3: positions 117-128 SEQ ID NO: 19, VL: CDR1: positions 47-58 SEQ ID NO: 23, CDR2: positions 76-78 SEQ ID NO: 23, CDR3: positions 115-123 SEQ ID NO: 23,

(vii) VH: CDR1: positions 45-53 SEQ ID NO: 19, CDR2: positions 71-78 SEQ ID NO: 19, CDR3: positions 117-128 SEQ ID NO: 19, VL: CDR1: positions 47 -58 SEQ ID NO: 26, CDR2: positions 76-78 SEQ ID NO: 26, CDR3: positions 115-123 SEQ ID NO: 26,

(viii) VH: CDR1: positions 45-53 SEQ ID NO: 19, CDR2: positions 71-78 SEQ ID NO: 19, CDR3: positions 117-128 SEQ ID NO: 19, VL: CDR1: positions 47 -58 SEQ ID NO: 27, CDR2: positions 76-78 SEQ ID NO: 27, CDR3: positions 115-123 SEQ ID NO: 27 and

(ix) VH: CDR1: positions 45-53 SEQ ID NO: 19, CDR2: positions 71-78 SEQ ID NO: 19, CDR3: positions 117-128 SEQ ID NO: 19, VL: CDR1: positions 47 -52 SEQ ID NO: 28, CDR2: positions 70-72 of SEQ ID NO: 28, CDR3: positions 109-117 of SEQ ID NO: 28.

In other preferred embodiments, an antibody having the ability to bind to CLDN18.2 preferably comprises one or more complementarity determining regions (CDRs), preferably at least a heavy chain variable region (VH) CDR3 and/or a light chain variable region (VL) an anti-CLDN18.2 monoclonal antibody, preferably an anti-CLDN18.2 monoclonal antibody described herein, and preferably contains one or more complementarity determining regions (CDRs), preferably at least the CDR3 variable region of the heavy chain variable regions (VH) and/or light chain variable regions (VL) described herein. In one embodiment, said one or more complementarity determining regions (CDRs) are selected from the set of complementarity determining regions CDR1, CDR2, and CDR3 described herein. In a particularly preferred embodiment, the antibody having the ability to bind to CLDN18.2 preferably comprises the CDR1, CDR2 and CDR3 regions of complementarity of the heavy chain variable region (VH) and/or the light chain variable region (VL) of an anti-CLDN18.2 monoclonal antibody, preferably a monoclonal antibody against CLDN18.2 described herein, and preferably contains CDR1, CDR2 and CDR3 complementarity regions of the heavy chain variable regions (VH) and/or light chain variable regions (VL) described herein.

In one embodiment, an antibody comprising one or more CDRs, a set of CDRs, or a combination of sets of CDRs as described herein contains said CDRs along with their intermediate framework regions. Preferably, this portion will also include at least about 50% of one or both of the first and fourth framework regions, with 50% being the C-terminal 50% of the first framework region and the N-terminal 50% of the fourth framework region. The construction of antibodies produced by recombinant DNA techniques may result in the introduction of N- or C-terminal residues into the variable regions encoded by linkers introduced to facilitate cloning or other manipulations, including the introduction of linkers to attach the variable regions of the invention to additional protein sequences, including heavy chains. immunoglobulin, other variable domains (for example, in the production of diabodies) or protein labels.

In one embodiment, an antibody containing one or more CDRs, a set of CDRs, or a combination of sets of CDRs, as described herein, contains said CDRs in a human antibody framework.

Reference herein to an antibody containing, relative to its heavy chain, a particular chain, or a particular region or sequence, preferably refers to the situation where all heavy chains of said antibody contain that particular chain, region, or sequence. This applies, respectively, to the light chain of an antibody.

It should be understood that the antibodies described herein can be delivered to a patient by administering a nucleic acid, such as an RNA encoding an antibody, and/or by administering a host cell containing a nucleic acid, such as an RNA encoding an antibody. Thus, the nucleic acid encoding the antibody, when administered to a patient, may be present in uncoated form or in a suitable delivery vehicle, such as in the form of liposomes or viral particles, or within a host cell. The provided nucleic acid can produce the antibody for long periods of time in a stable manner, reducing the instability, at least in part, seen with therapeutic antibodies. Nucleic acids to be delivered to a patient may be produced by recombinant means. If the nucleic acid is administered to a patient without being present in a host cell, it is preferably taken up by the patient's cells to express the antibody encoded by the nucleic acid. If the nucleic acid is administered to a patient in a host cell, it is preferably expressed by the host cell within the patient to produce an antibody encoded by the nucleic acid.

As used herein, the term "nucleic acid" is intended to include DNA and RNA such as genomic DNA, cDNA, mRNA, recombinantly produced and chemically synthesized molecules. The nucleic acid may be single-stranded or double-stranded. RNA includes in vitro transcribed RNA (IVT RNA) or synthetic RNA.

Nucleic acids may be included in the vector. As used herein, the term "vector" includes any vectors known to those skilled in the art, including plasmid vectors, cosmid vectors, phage vectors such as lambda phage, viral vectors such as adenovirus or baculovirus vectors, or artificial chromosome vectors, such as bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC) or P1 artificial chromosomes (PAC). These vectors are expression vectors as well as cloning vectors. Expression vectors contain plasmids as well as viral vectors and typically contain the coding sequence of interest and the corresponding DNA sequences necessary for expression of the operably linked coding sequence in a particular host organism (e.g., bacteria, yeast, plant, insect, or mammal) or in expression systems in vitro. Cloning vectors are typically used to process and amplify a particular DNA fragment of interest and may not have the functional sequences necessary to express the DNA fragments of interest.

In the context of the present invention, the term "RNA" refers to a molecule that contains ribonucleotide residues and preferably consists entirely or essentially of ribonucleotide residues. "Ribonucleotide" refers to a nucleotide with a hydroxyl group at the 2' position of the β-D-ribofuranosyl group. The term includes double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, substantially purified RNA, synthetic RNA, recombinantly produced RNA, and modified RNA that differs from natural RNA by the addition, deletion, substitution and/or substitution of one or several nucleotides. Such changes may include the addition of non-nucleotide material, for example, to the end(s) of the RNA or inward, for example, in one or more nucleotides of the RNA. Nucleotides in RNA molecules may also contain non-standard nucleotides such as non-natural nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs may be referred to as natural RNA analogs or analogs.

According to the present invention, the term "RNA" includes and preferably refers to "mRNA", which means "messenger RNA" and refers to a "transcript" that can be obtained using DNA as a template and which encodes a peptide or protein. An mRNA typically includes a 5' untranslated region (5'UTR), a protein or peptide coding region, and a 3' untranslated region (3'UTR). mRNA has a limited half-life in cells and in vitro. Preferably, mRNA is obtained by in vitro transcription using a DNA template. In one embodiment of the invention, RNA is obtained by in vitro transcription or chemical synthesis. The methodology for in vitro transcription is known to those skilled in the art. For example, there are many in vitro transcription kits commercially available.

In order to increase the expression and/or stability of the RNA used according to the present invention, it can be modified, preferably without changing the sequence of the expressed peptide or protein.

The term "modification" in the context of RNA, which is used according to the present invention, includes any modification of the RNA, which is not naturally found in the specified RNA. Such modified RNA is encompassed herein by the term "RNA".

For example, the RNA according to the invention may have modified natural or synthetic ribonucleotides to increase its stability and/or reduce its cytotoxicity. For example, in one embodiment, in the RNA used according to the invention, 5-methylcytidine is substituted partially or completely, preferably completely, for cytidine. Alternatively or additionally, in one embodiment, in the RNA used according to the invention, pseudouridine replaces partially or completely, preferably completely, uridine.

In one embodiment, the term "modification" refers to providing the RNA with a 5'-cap structure or an analog of the 5'-cap structure. The term "5' cap" refers to the cap structure found at the 5' end of the mRNA molecule and usually consists of a guanosine nucleotide linked to the mRNA via an unusual 5'-5' triphosphate bond. In one embodiment, this guanosine is methylated at the 7-position. The term "traditional 5'-cap" refers to a naturally occurring RNA 5'-cap, preferably a 7-methylguanosine (m7G) cap. In the context of the present invention, the term "5'-cap" includes a 5'-cap analog that resembles the structure of an RNA cap and is modified to have the ability to stabilize RNA when attached to it, preferably in vivo and/or in a cell.

Preferably, the RNA, if delivered, i.e. is transfected into a cell, in particular the cell present in vivo has expressed the protein or peptide it encodes.

The term "transfection" refers to the introduction of nucleic acids, specifically RNA, into a cell. For the purposes of the present invention, the term "transfection" also includes the introduction of a nucleic acid into a cell, or the absorption of a nucleic acid into such a cell, in which the cell may be present in an object, such as a patient. Thus, according to the present invention, the nucleic acid transfection cell described herein may be present in vitro or in vivo, for example, the cell may form part of an organ, tissue and/or body of a patient. According to the invention, transfection may be transient or stable. For some applications of transfection, it is sufficient that the transfected genetic material is only transiently expressed. Since the nucleic acid introduced into the transfection process usually does not integrate into the nuclear genome, the foreign nucleic acid will be diluted during mitosis or degraded. Cells that allow episomal amplification of nucleic acids significantly reduce the degree of dilution. If it is desired that the transfected nucleic acid actually remain in the cell's genome and in its daughter cells, then stable transfection must take place. The RNA can be transfected into cells for transient expression of the encoded protein.

The term "stability" of RNA refers to the "half-life" of RNA. "Half-life" refers to the period of time that is required to eliminate half of the activity, number or number of molecules. In the context of the present invention, the half-life of an RNA is indicative of the stability of said RNA. The half-life of RNA can influence the "length of expression" of the RNA. An RNA having a long half-life can be expected to be expressed over a long period of time.

In the context of the present invention, the term "transcription" refers to the process in which the genetic code in a DNA sequence is transcribed into RNA. Subsequently, RNA can be translated into protein. According to the present invention, the term "transcription" includes "in vitro transcription", where the term "in vitro transcription" refers to a method in which RNA, specifically mRNA, is synthesized in vitro in a cell-free system, preferably using suitable cell extracts. Preferably, cloning vectors are used to generate transcripts. These cloning vectors are commonly referred to as transcription vectors and are encompassed by the term "vector" according to the present invention.

The term "translation" according to the invention refers to the process in the ribosomes of a cell by which a messenger RNA chain directs the assembly of an amino acid sequence to produce a peptide or protein.

The term "expression" is used according to the invention in its most general sense and includes the production of RNA and/or peptides or proteins, for example, by transcription and/or translation. With regard to RNA, the term "expression" or "translation" refers in particular to the formation of peptides or proteins. It also includes partial expression of nucleic acids. Moreover, expression can be transient or stable. According to the invention, the term expression also includes "aberrant expression" or "abnormal expression".

"Aberrant expression" or "abnormal expression" means according to the invention that the expression is changed, preferably increased, compared to the reference, for example, when the subject does not have a disease associated with aberrant or abnormal expression of a certain protein, for example, a tumor antigen. An increase in expression refers to an increase of at least 10%, specifically at least 20%, at least 50%, or at least 100% or more. In one embodiment, expression is found only in diseased tissue, while expression in healthy tissue is repressed.

The term "specifically expressed" means that the protein is essentially expressed only in a particular tissue or organ. For example, a tumor antigen specifically expressed in the gastric mucosa means that said protein is predominantly expressed in the gastric mucosa and is not expressed in other tissues or is not expressed to a significant extent in other tissues or organs. Thus, a protein that is exclusively expressed in gastric mucosal cells and to a much lesser extent in any other tissue, such as the testis, will be specifically expressed in gastric mucosal cells. In some embodiments, the tumor antigen may also be specifically expressed under normal conditions in more than one tissue or organ type, such as 2 or 3 tissue or organ types, but preferably no more than 3 different tissue or organ types. In this case, the tumor antigen is then specifically expressed in these organs. For example, if a tumor antigen is expressed under normal conditions, preferably approximately equally in the lungs and stomach, then said tumor antigen is specifically expressed in the lungs and stomach.

According to the invention, the term "coding RNA" means that the RNA, if present in an appropriate environment, preferably within a cell, can be expressed to produce the protein or peptide it encodes.

Some aspects of the invention are based on the adoptive transfer of host cells that are transfected in vitro with a nucleic acid, such as an RNA encoding an antibody described herein, and transferred to recipients, such as patients, preferably after ex vivo production, ranging from low represented progenitors to clinically significant numbers of cells. The host cells used for the treatment according to the invention may be autologous, allogeneic, or syngeneic to the treated recipient.

The term "autologous" is used to describe everything that comes from the same entity. For example, "autologous transplantation" refers to transplantation of tissue or organs derived from the same subject. Such treatments are beneficial because they overcome the immunological barrier that would otherwise lead to rejection.

The term "allogeneic" is used to describe anything that comes from different individuals of the same species. Two or more individuals are said to be allogeneic to each other when the genes at one or more loci are not identical.

The term "syngeneic" is used to describe everything that comes from individuals or tissues that have identical genotypes, i.e. identical twins or animals of the same inbred strain or their tissues.

The term "heterologous" is used to describe something that is made up of several different elements. As an example, the transfer of bone marrow from one person to another person is a heterologous transplant. A heterologous gene is a gene derived from a source other than the subject.

The term "peptide" according to the invention includes oligo- and polypeptides and refers to substances containing two or more, preferably 3 or more, preferably 4 or more, preferably 6 or more, preferably 8 or more, preferably 9 or more, preferably 10 or more , preferably 13 or more, preferably another 16, preferably 21 or more and preferably up to 8, 10, 20, 30, 40 or 50, in particular 100 amino acids covalently linked by peptide bonds. The term "protein" refers to large peptides, preferably peptides with more than 100 amino acid residues, but in general the terms "peptides" and "proteins" are synonymous and are used interchangeably herein.

Guidance given herein with respect to specific amino acid sequences, such as those provided in a sequence listing, should be construed to also refer to variants of said specific sequences, resulting in sequences that are functionally equivalent to said specific sequences, such as amino acid sequences, having properties identical or similar to those of specific amino acid sequences.

One of the important properties is to maintain the binding of the antibody to its target or to maintain the effector functions of the antibody. Preferably, a sequence that is variant with respect to a particular sequence, when replaced by a particular sequence in an antibody, retains the binding of said antibody to its target and preferably the functions of said antibody as described herein, e.g., CDC-mediated lysis or ADCC-mediated lysis .

Those skilled in the art will appreciate that, in particular, CDR sequences, hypervariable and variable regions can be modified without loss of the ability of an antibody to bind to its target. For example, the CDR regions will either be identical to or highly homologous to the regions of the antibodies referred to herein. By "highly homologous" is meant that 1 to 5 substitutions, preferably 1 to 4, eg 1 to 3 or 1 or 2 substitutions can be made in the CDR. In addition, hypervariable and variable regions can be modified so that they show significant homology to the antibody regions specifically disclosed herein.

The term "variant" according to the invention refers in particular to mutants, splicing variants, conformations, isoforms, allelic variants, species and species homologues, in particular to those found in nature. An allelic variant refers to a change in the normal sequence of a gene, the meaning of which is often unclear. Whole gene sequencing will often identify multiple allelic variants for a given gene. A species homologue is a nucleic acid or amino acid sequence of a species of other origin derived from a given nucleic acid sequence or amino acid sequence. The term "variant" shall encompass any post-translationally modified variants and conformational variants.

For the purposes of the present invention, amino acid sequence "variants" include amino acid insertion variants, amino acid addition variants, amino acid deletion variants, and/or amino acid substitution variants. Amino acid deletion variants that include deletion at the N-terminus and/or C-terminus of the protein are also referred to as N-terminal and/or C-terminal truncated variants.

Amino acid insertion variants include insertions of one or two or more amino acids in a particular amino acid sequence. In the case of amino acid sequence variants having an insert, one or more amino acid residues are introduced at a specific site in the amino acid sequence, although random insertion with appropriate screening of the resulting product is also possible.

Options for adding amino acids include amino and/or carboxy-terminal fusions of one or more amino acids, for example, 1, 2, 3, 5, 10, 20, 30, 50 or more amino acids.

Amino acid deletion variants are characterized by the removal of one or more amino acids from a sequence, for example, the removal of 1, 2, 3, 5, 10, 20, 30, 50 or more amino acids. Deletions can be in any position of the protein.

Amino acid variants are characterized by at least one residue in the deleted sequence and another residue inserted in its place. Preference is given to modifications that are at positions in the amino acid sequence that are not conservative between homologous proteins or peptides and/or substitutions of amino acids for others having similar properties. Preferably, amino acid substitutions in protein variants are conservative amino acid substitutions, that is, substitutions of equally charged or uncharged amino acids. A conservative amino acid substitution includes a substitution with one from a family of amino acids that are related in their side chains. Natural amino acids are usually divided into four families: acidic (aspartate, glutamate), basic (lysine, arginine, histidine), non-polar (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan) and uncharged polar (glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine) amino acids. Phenylalanine, tryptophan, and tyrosine are sometimes classified as aromatic amino acids.

Preferably, the degree of similarity, preferably identity, between a given amino acid sequence and an amino acid sequence that is a variant of said amino acid sequence will be at least about 60%, 65%, 70%, 80%, 81%, 82%, 83% , 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. The degree of similarity or identity is given preferably for an amino acid region that is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or about 100% of the entire length of the reference amino acid sequence. For example, if the reference amino acid sequence is 200 amino acids, the degree of similarity or identity is given preferably for at least about 20, at least about 40, at least about 60, at least about 80, at least about 100, at least about 120, at least about 140, at least about 160, at least about 180, or about 200 amino acids, preferably continuous amino acids. In preferred embodiments, the degree of similarity or identity is given for the entire length of the reference amino acid sequence. Alignment to determine sequence similarity, preferably sequence identity, can be done with known tools, preferably using the best sequence alignment, e.g. using Align, using standard settings, preferably EMBOSS::needle, Matrix: Blosum62, gap opening penalty 10.0, penalty for gap extension 0.5.

"Sequence similarity" refers to the percentage of amino acids that are either identical or are conservative amino acid substitutions. "Sequence identification" between two amino acid sequences refers to the percentage of amino acids that are identical between sequences.

The term "percent identity" is intended to mean the percentage of amino acid residues that are identical between the two sequences to be compared, obtained after the best alignment, this percentage being purely statistical, and the differences between the two sequences being distributed randomly and along the entire length. Sequence comparisons between two amino acid sequences are typically performed by comparing those sequences after they are optimally aligned, said comparison being made using a segment or "comparison window" to identify and compare local areas of sequence similarity. The optimal sequence alignment for comparison can also be obtained manually using the local homology algorithm of Smith and Waterman, 1981, Ads App. Math. 2, 482, using the local homology algorithm of Neddleman and Wunsch, 1970, J. Mol. Biol. 48, 443 using the similarity search method Pearson and Lipman, 1988, Proc. Natl. Acad. sci. USA 85, 2444 or by computer programs that use these algorithms (GAP, BESTFIT, FASTA, BLAST P, BLAST N, and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Drive, Madison, Wisconsin).

Percent identity is calculated by determining the number of identical positions between two compared sequences, dividing this number by the number of compared positions and multiplying the result by 100 to obtain the percentage identity between the two sequences.

The term "cell" or "host cell" preferably refers to an intact cell, that is, a cell with an intact membrane that does not release its normal intracellular components such as enzymes, organelles, or genetic material. The intact cell is preferably a viable cell, ie a living cell capable of performing its normal metabolic functions. Preferably, said term refers according to the invention to any cell that can be transfected with exogenous nucleic acid. Preferably, a cell transfected with an exogenous nucleic acid and transferred to a recipient can express the nucleic acid in the recipient. The term "cell" includes bacterial cells; other useful cells are yeast cells, fungal cells or mammalian cells. Suitable bacterial cells include those from Gram-negative bacterial strains such as Escherichia coli, Proteus and Pseudomonas strains and Gram-positive bacterial strains such as Bacillus, Streptomyces, Staphylococcus and Lactococcus strains. Suitable fungal cells include cells from Trichoderma, Neurospora and Aspergillus species. Suitable yeast cells include cells from Saccharomyces species (eg Saccharomyces cerevisiae), Schizosaccharomyces (eg Schizo saccharomyces pombe), Pichia (eg Pichia pastoris and Pichia methanolica) and Hansenula. Suitable mammalian cells include, for example, CHO cells, BHK cells, HeLa cells, COS cells, 293 HEK, and the like. However, amphibian cells, insect cells, plant cells, and any other cells used in the art for expressing heterologous proteins can also be used. Mammalian cells are particularly preferred for adoptive transfer, such as those of humans, mice, hamsters, pigs, goats, and primates. Cells can be obtained from a wide variety of tissue types and include primary cells and cell lines such as cells of the immune system, in particular antigen presenting cells such as dendritic cells and T cells, stem cells such as hematopoietic stem cells and mesenchymal stem cells and other types of cells. An antigen presenting cell is a cell that presents an antigen in the context of a major histocompatibility complex on its surface. T cells can recognize this complex using their T cell receptor (TCR).

The term "transgenic animal" refers to an animal having a genome containing one or more transgenes, preferably heavy and/or light chain transgenes, or transchromosomes (either integrated or not integrated into the animal's natural genomic DNA) and which is preferably capable of expressing the transgenes. For example, a transgenic mouse may have a human light chain transgene and either a human heavy chain transgene or a human heavy chain transchromosome such that the mouse produces antibodies against a human tumor antigen when immunized with the tumor antigen and/or cells expressing the tumor antigen. The human heavy chain transgene can be integrated into mouse chromosomal DNA, as is the case for transgenic mice, e.g., HuMAb mice, such as HCo7 or HCol2 mice, or the human heavy chain transgene can be maintained extrachromosomally, as is the case for transchromosomal (e.g., KM) mice , as described in WO 02/43478. Such transgenic and transchromosomal mice may be capable of producing multiple isotypes of human monoclonal antibodies to a tumor antigen (eg, IgG, IgA, and/or IgE) by VDJ recombination and isotype switching.

"Reduce", "reduce", or "inhibit", as used herein, means an overall reduction or the ability to cause an overall reduction, preferably 5% or more, 10% or more, 20% or more, more preferably 50% % or more, and most preferably 75% or more, of a level, such as an expression level or a cell proliferation level.

Terms such as "increase" or "increase" preferably refer to an increase or increase of about 10%, preferably at least 20%, preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 80%, and most preferably at least 100%, at least 200%, at least by 500%, at least 1000%, at least 10000% or even more.

The antibodies described herein can be prepared in a variety of ways, including conventional monoclonal antibody methodology, such as the standard somatic cell hybridization technique of Kohler and Milstein, Nature 256:495 (1975). Although somatic cell hybridization procedures are preferred, other methods for producing monoclonal antibodies can in principle be used, such as viral or oncogenic transformation of B-lymphocytes, or phage display methods using antibody gene libraries.

The preferred animal system for producing hybridomas that secrete monoclonal antibodies is the murine system. The production of hybridoma in mice is a very well established procedure. Immunization protocols and methods for isolating immunized splenocytes for fusion are known in the art. Fusion partners (eg, murine myeloma cells) and fusion procedures are also known.

Other preferred animal systems for producing hybridomas that secrete monoclonal antibodies are the rat and rabbit system (e.g., described in Spieker-Polet et al., Proc. Natl. Acad. Sci. U.S.A. 92:9348 (1995), see also Rossi et al., Am J Clin Pathol 124: 295 (2005)).

In yet another preferred embodiment, human monoclonal antibodies can be generated using transgenic or transchromosomal mice carrying parts of the human immune system rather than the mouse system. These transgenic and transchromosomal mice include the mice known as HuMAb mice and KM mice, respectively, and are collectively referred to herein as "transgenic mice". Production of human antibodies in such transgenic mice can be performed as detailed for CD20 in WO2004035607.

Another strategy for obtaining monoclonal antibodies is to directly isolate genes encoding antibodies from lymphocytes that produce antibodies of a certain specificity, for example, see Babcock et al., 1996; A new strategy for obtaining monoclonal antibodies from single isolated lymphocytes producing antibodies of certain specificities. For details on the development of recombinant antibodies, see also Welschof and Kraus, Recombinant antibodes for cancer therapy ISBN-0-89603-918-8 and Benny K.C. Lo Antibody Engineering ISBN 1-58829-092-1.

To generate antibodies, mice can be immunized with carrier-conjugated peptides derived from the sequence of the antigen, i.e., the sequence against which antibodies of an enriched preparation of recombinantly expressed antigen or fragments thereof and/or cells expressing the antigen are to be directed, as described. Alternatively, mice can be immunized with DNA encoding the antigen or fragments thereof. In the event that immunization with a purified or enriched antigen preparation does not produce antibodies, mice can also be immunized with antigen expressing cells, such as a cell line, to stimulate immune responses.

The immune response can be monitored during the course of immunization with plasma and serum samples obtained from the tail vein or by retroorbital phlebotomy. Mice with sufficient immunoglobulin titer can be used for fusion. Mice can be stimulated ip or iv with antigen expressing cells 3 days prior to sacrifice and spleen removal to increase the percentage of hybridomas secreting specific antibodies.

To obtain monoclonal antibody-producing hybridomas, splenocytes and lymph node cells can be isolated from immunized mice and fused to an appropriate immortalized cell line, such as a mouse myeloma cell line. The resulting hybridomas can then be screened for antigen-specific antibodies. Individual wells can then be screened by ELISA for antibody-secreting hybridomas. When analyzing immunofluorescence and FACS using cells expressing the antigen, it is possible to identify antibodies with specificity for the antigen. Antibody-secreting hybridomas can be replicated, screened again and, if still positive for monoclonal antibodies, can be subcloned by the limiting dilution method. The stable subclones can then be cultured in vitro to generate antibodies in tissue culture media for characterization.

Antibodies can also be generated in the transfectome of a host cell using, for example, a combination of recombinant DNA techniques and gene transfection techniques that are well known in the art (Morrison, S. (1985) Science 229: 1202).

For example, in one embodiment, the gene(s) of interest, such as antibody genes, can be ligated into an expression vector, such as a eukaryotic expression plasmid, such as used by the GS gene expression system disclosed in WO 87/04462, WO 89/01036 and EP 338 841 or other expression systems well known in the art. The purified plasmid with cloned antibody genes can be introduced into eukaryotic host cells such as CHO cells, NS/0 cells, HEK293T cells or HEK293 cells, or alternatively other eukaryotic cells such as plant-derived, fungal or yeast cells. The method used to introduce these genes may be a method described in the art, such as electroporation, the use of lipofectin, lipofectamine, or others. Once these antibody genes are introduced into host cells, cells expressing the antibody can be identified and selected. These cells are transfectomes which can then be amplified for their level of expression and worked up to produce antibodies. Recombinant antibodies can be isolated and purified from these culture supernatants and/or cells.

Alternatively, cloned antibody genes may be expressed in other expression systems, including prokaryotic cells such as microorganisms such as E. coli. In addition, antibodies can be produced in non-human transgenic animals, such as sheep and rabbit milk or chicken eggs, or transgenic plants; See, for example, Verma, R., et al. (1998) J. Immunol. Meth. 216:165-181; Pollock, et al. (1999) J. Immunol. Meth. 231:147-157; and Fischer, R., et al. (1999) Biol. Chem. 380: 825-839.

Chimerization

Mouse antibodies are highly immunogenic in humans upon repeated use, resulting in a reduced therapeutic effect. The main immunogenicity is mediated by heavy chain constant regions. The immunogenicity of mouse antibodies in humans can be reduced or completely eliminated if the corresponding antibodies are chimerized or humanized. Chimeric antibodies are antibodies whose different parts are derived from different animal species, for example, which have a variable region derived from a mouse antibody and human immunoglobulin constant regions. Antibody chimerization is achieved by combining the heavy and light chain variable regions of a mouse antibody with a human heavy and light chain constant region (e.g., as described by Kraus et al., in Methods in Molecular Biology series, Recombinant antibodies for cancer therapy ISBN-0-89603-918 -eight). In a preferred embodiment, the chimeric antibodies are generated by fusing a human kappa light chain constant region with a mouse light chain variable region. In a further preferred embodiment, chimeric antibodies can be generated by fusing a human lambda light chain constant region with a mouse light chain variable region. Preferred heavy chain constant regions for making chimeric antibodies are IgG1, IgG3 and IgG4. Other preferred heavy chain constant regions for making chimeric antibodies are IgG2, IgA, IgD and IgM.

humanization

Antibodies interact with target antigens primarily through amino acid residues that are located in the six complementarity-determining regions of the heavy and light chains (CDRs). For this reason, the amino acid sequences within the CDR are more diverse between individual antibodies than those outside the CDR. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from a specific naturally occurring antibody grafted onto framework sequences from another antibody with other properties (see, for example, Riechmann, L. et al. (1998) Nature 332: 323-327; Jones, P. et al. (1986) Nature 321: 522-525; and Queen, C. et al. (1989) Proc. Natl. Acad. Sci. U. S. A. 86: 10029-10033). Such framework sequences can be obtained from public DNA databases that include germline antibody gene sequences. These germline sequences will differ from mature antibody gene sequences because they will not include the fully assembled variable genes that are generated by V(D)J fusion during B cell maturation. The germline gene sequences will also differ from high affinity secondary repertoire antibody sequences when individually evenly distributed across the variable region.

The ability of antibodies to bind an antigen can be determined using standard binding assays (eg, ELISA, Western blot, immunofluorescence, and flow cytometric analysis).

For antibody purification, selected hybridomas can be grown in 2 liter spinner flasks for monoclonal antibody purification. Otherwise, antibodies can be produced in bioreactors based on dialysis. Supernatants can be filtered and, if necessary, concentrated prior to affinity chromatography on either Protein G Sepharose or Protein A Sepharose. Eluted IgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity. The buffer solution can be replaced with PBS and the concentration can be determined by OD280 using an absorbance of 1.43. Monoclonal antibodies can be divided into aliquots and stored at -80°C.

In order to determine whether selected monoclonal antibodies can bind to unique epitopes, site-directed or multisite-directed mutagenesis can be used.

To determine the isotype of antibodies, isotype ELISAs can be performed with various commercial kits (eg, Zymed, Roche Diagnostics). Microplate wells can be coated with anti-mouse protein Ig. After blocking, the plates are reacted with monoclonal antibodies or purified isotype controls at ambient temperature for two hours. The contents of the wells can then be reacted with IgG1, IgG2a, IgG2b or IgG3, IgA, or mouse IgM-specific peroxidase-conjugated probes. After washing, plates can be developed with ABTS substrate (1 mg/ml) and analyzed at OD 405-650. Alternatively, the IsoStrip Mouse Monoclonal Antibody Isotyping Kit (Roche, cat. no. 1493027) can be used as described by the manufacturer.

Flow cytometry can be used to demonstrate the presence of antibodies in the sera of immunized mice or the binding of monoclonal antibodies to living cells expressing the antigen. Cell lines naturally or transfected expressing antigen and negative controls lacking antigen expression (grown under standard growth conditions) can be mixed with various concentrations of monoclonal antibodies in hybridoma supernatants or in PBS containing 1% FBS and can be incubated at 4°C for 30 minutes. After washing, the APC- or Alexa647-labeled anti-IgG antibody can bind to the antigen-bound monoclonal antibody under the same conditions as the primary antibody staining conditions. Samples can be analyzed by flow cytometry using a FACS instrument using the properties of light and side scatter with the transmission of single live cells. To distinguish antigen-specific monoclonal antibodies from non-specific binders in a single measurement, a co-transfection method can be used. Cells transiently transfected with plasmids encoding an antigen and a fluorescent marker can be stained as described above. Transfected cells can be detected in a different fluorescence channel than cells stained with antibodies. Because most transfected cells express both transgenes, antigen-specific monoclonal antibodies bind preferentially to cells expressing the fluorescence marker, while non-specific antibodies bind in comparable proportions to non-transfected cells. An alternative analysis using fluorescence microscopy can be used in addition to or instead of flow cytometry analysis. Cells can be stained exactly as described above and examined using fluorescence microscopy.

To demonstrate the presence of antibodies in the sera of immunized mice, or the binding of monoclonal antibodies to living cells expressing the antigen, immunofluorescence microscopy assay can be used. For example, cell lines expressing either spontaneously or after transfection the antigen and negative controls lacking antigen expression are grown in chamber slides under standard growth conditions in DMEM/F12 medium supplemented with 10% fetal calf serum (FCS), 2 mM L- glutamine, 100 IU/ml penicillin, and 100 µg/ml streptomycin. The cells can then be fixed with methanol or paraformaldehyde or left untreated. The cells can then react with monoclonal antibodies against the antigen within 30 minutes. at 25°C. After washing, cells can react with Alexa555-labeled anti-mouse IgG antibody (Molecular Probes) under the same conditions. The cells can then be examined using fluorescence microscopy.

Cell extracts from cells expressing the antigen and corresponding negative controls can be prepared and subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. After electrophoresis, the separated antigens are transferred to nitrocellulose membranes, blocked, and examined for the monoclonal antibodies to be tested. IgG binding can be detected using anti-mouse protein IgG peroxidase and developed with an ECL substrate.

Antibodies can be further tested for reactivity with the antigen by an immunohistochemistry method well known to the skilled artisan, such as using paraformaldehyde or acetone fixed cryosections or paraformaldehyde-fixed paraffin-embedded sections from non-neoplastic or tumor tissue samples obtained from patients during a routine surgical procedure. , or from mice bearing xenotransformed tumors inoculated with cell lines expressing the antigen spontaneously or after transfection. according to the supplier's instructions.

Antibodies can be tested for their ability to mediate phagocytosis and killing of cells expressing the tumor antigen. In vitro testing of monoclonal antibody activity will provide an initial screen before testing in vivo models.

Antibody-dependent cell-mediated cytotoxicity (ADCC)

Briefly, polymorphonuclear cells (PMN), NK cells, monocytes, mononuclear cells or other effector cells from healthy donors can be purified by Ficoll Hypaque centrifugation followed by lysis of contaminating red blood cells. Washed effector cells can be suspended in RPMI supplemented with 10% heat-inactivated fetal calf serum, or alternatively with 5% inactivated human serum, and mixed with ⁵¹ Cr-labelled target cells expressing tumor antigen at different ratios of effector cells to cells. -targets. Alternatively, target cells can be labeled with a fluorescence enhancing ligand (BATDA). The highly fluorescent ligand-enhancing europium chelate that is released from dead cells can be measured with a fluorometer. Another alternative method may use transfection of target cells with luciferase. The added lucifer yellow can then only be oxidized by viable cells. Purified IgGs against tumor antigens can then be added at various concentrations. Extraneous human IgG can be used as a negative control. Assays can be performed within 4-20 hours at 37°C, depending on the type of effector cells used. Samples can be analyzed for cytolysis by measuring the release of ⁵¹ Cr or the presence of EuTDA chelate in the culture supernatant. Otherwise, the luminescence resulting from the oxidation of lucifer yellow may be a measure of viable cells.

Monoclonal antibodies against a tumor antigen can also be tested in various combinations to determine if cytolysis is enhanced using multiple monoclonal antibodies.

Complementary dependent cytotoxicity (CDC)

Monoclonal antibodies of tumor antigens can be tested for their ability to mediate CDC using a variety of known methods. For example, serum for complement can be obtained from blood in a manner known to those skilled in the art. Various methods can be used to determine CDC mAb activity. For example, ⁵¹ Cr release, or increased membrane permeability, can be measured using a propidium iodide (PI) exclusion assay. Briefly, target cells can be washed and 5×10 ⁵ /ml can be incubated with various concentrations of mAb for 10-30 minutes. At room temperature or at 37°C. Serum or plasma can then be added to a final concentration of 20% (v/v) and the cells are incubated at 37°C for 20-30 min. All cells from each sample can be added to the PI solution in the FACS tube. The mixture can then be immediately analyzed by flow cytometry analysis using the FACSArray.

In an alternative assay, CDC induction can be determined on adherent cells. In one embodiment of this assay, cells are seeded 24 hours prior to assay at a density of 3×10 ⁴ /well in culture flat-bottomed microplates. The next day, the growth medium is removed and the cells are incubated in triplicate with antibodies. Control cells are incubated with culture medium or growth medium containing 0.2% saponin to determine background lysis and maximum lysis, respectively. After incubation for 20 min at room temperature, the supernatant is removed and 20% (v/v) human plasma or serum in DMEM (preheated to 37°C) is added to the cells and incubated for another 20 min at 37°C. All cells from each sample are added to a solution of propidium iodide (10 μg/ml). The supernatants are then replaced with PBS containing 2.5 μg/ml ethidium bromide and the fluorescence emission upon excitation at 520 nm is measured at 600 nm using Tecan Safire. Percent specific lysis is calculated as follows: percent specific lysis = (background fluorescence fluorescence)/(fluorescence maximum background fluorescence fluorescence) × 100.

Apoptosis Induction and Cell Proliferation Inhibition by Monoclonal Antibodies

To test the ability to initiate apoptosis, monoclonal antibodies to tumor antigens can, for example, be incubated with tumor cells positive for tumor antigens or tumor cells transfected with tumor antigen at 37° C. for about 20 hours. Cells can be harvested, washed in annexin V binding buffer (BD biosciences) and incubated with FITC or APC conjugated annexin V (BD biosciences) for 15 min in the dark. All cells from each sample can be added to a solution of PI (10 μg/ml in PBS) in a FACS tube and immediately evaluated by flow cytometry (as above). Alternatively, overall inhibition of cell proliferation by monoclonal antibodies can be detected with commercially available kits. The DELFIA Cell Proliferation Kit (Perkin-Elmer, Cat. No. AD0200) is a non-isotopic immunoassay based on the measurement of 5-bromo-2'-deoxyuridine (BrdU) incorporation during DNA synthesis of proliferating cells in microplates. The incorporated BrdU is detected using a europium labeled monoclonal antibody. To ensure antibody detection, the cells are fixed and denatured with DNA using a Fix solution. The unbound antibody is washed off and the DELFIA inducer is added to dissociate the europium ions from the labeled antibody into solution where they form highly fluorescent chelates with the components of the DELFIA inducer. Measured fluorescence—using time-resolved fluorometry in detection—is proportional to DNA synthesis in each well well.

Preclinical studies

The antibodies described herein can also be tested in an in vivo model (e.g., immunodeficient mice bearing tumor xenografts inoculated with tumor antigen expressing cell lines to determine their effectiveness in controlling the growth of tumor antigen expressing tumor cells.

In vivo studies following xenotransplantation of tumor cells expressing a tumor antigen into immunocompromised mice or other animals can be performed using the antibodies described herein. Antibodies can be administered to mice without tumors followed by injection of tumor cells to measure the effects of the antibodies in preventing tumor formation or tumor-related symptoms. Antibodies can be administered to tumor-bearing mice to determine the therapeutic efficacy of the respective antibodies in reducing tumor growth, metastases, or tumor-related symptoms. The use of antibodies can be combined with the use of other substances as cystostatic drugs, growth factor inhibitors, cell cycle blockers, angiogenesis inhibitors, or other antibodies to determine the synergistic efficacy and potential toxicity of the combinations. For analysis of antibody-mediated toxic side effects, animals can be inoculated with antibodies or control reagents and closely examined for symptoms that may be associated with tumor antigen therapy. Possible side effects of the use of antibodies against tumor antigens in vivo, in particular, include toxicity in tissues expressing tumor antigens. Antibodies that recognize tumor antigen in humans and other species such as mice are particularly useful in predicting potential side effects mediated by the use of monoclonal antibodies to tumor antigens in humans.

Mapping of epitopes recognized by antibodies can be performed as detailed in "Epitope Mapping Protocols (Methods in Molecular Biology)" by Glenn E. Morris ISBN-089603-375-9 and in "Epitope Mapping: A Practical Approach" Practical Approach Series, 248 by Olwyn M. R. Westwood, Frank C. Hay.

The compounds and agents described herein may be administered in the form of any suitable pharmaceutical composition.

Pharmaceutical compositions are preferably sterile and contain an effective amount of the antibodies described herein, and optionally additional agents discussed herein, to obtain the desired response or desired effect.

Pharmaceutical compositions are usually provided in a single dosage form and can be prepared in a known manner. The pharmaceutical composition may, for example, be in the form of a solution or suspension.

The pharmaceutical composition may contain salts, buffers, preservatives, carriers, diluents and/or excipients, each of which is preferably pharmaceutically acceptable. The term "pharmaceutically acceptable" refers to the non-toxicity of a material that does not interact with the action of the active ingredient of the pharmaceutical composition.

Salts that are not pharmaceutically acceptable can be used to prepare pharmaceutically acceptable salts and are included in the invention. Pharmaceutically acceptable salts of this type include, without limitation, those derived from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic acids, and the like. Pharmaceutically acceptable salts may also be prepared as alkali metal salts or alkaline earth metal salts such as sodium salts, potassium salts or calcium salts.

Suitable buffer substances for use in the pharmaceutical composition include acetic acid in the salt, citric acid in the salt, boric acid in the salt, and phosphoric acid in the salt.

Suitable preservatives for use in the pharmaceutical composition include benzalkonium chloride, chlorobutanol, paraben and thimerosal.

The injectable formulation may contain a pharmaceutically acceptable excipient such as Ringer's lactate.

The term "carrier" refers to an organic or inorganic component of a natural or synthetic nature, in which the active component is combined to facilitate, enhance or use. According to the invention, the term "carrier" also includes one or more compatible solid or liquid excipients, diluents, or encapsulating agents that are suitable for administration to a patient.

Possible carrier substances for parenteral administration are, for example, sterile water, Ringer's solution, Ringer's lactate, sterile sodium chloride solution, polyalkylene glycols, hydrogenated naphthalenes and in particular biocompatible lactide polymers, lactide-glycolide copolymers or polyoxyethylene/polyoxypropylene copolymers.

As used herein, the term "excipient" is intended to refer to all substances that may be present in a pharmaceutical composition that are not active ingredients, such as, for example, carriers, binders, lubricants, thickeners, surfactants, preservatives, emulsifiers. , buffers, flavors or colors.

The agents and compositions described herein may be administered by any conventional route, such as parenteral administration, including injection or infusion. Administration is preferably parenteral, eg intravenous, intraarterial, subcutaneous, intradermal or intramuscular.

Compositions suitable for parenteral administration generally contain a sterile aqueous or non-aqueous preparation of the active compound, which is preferably isotonic with the recipient's blood. Examples of compatible vehicles and solvents are Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fatty oils are usually used as a solution or suspension medium.

The agents and compositions described herein are administered in effective amounts. "Effective amount" refers to the amount that achieves the desired response or desired effect alone or together with additional doses. In the case of treating a specific disease or a specific condition, the desired response preferably refers to the inhibition of the course of the disease. This includes slowing down the progress of the disease, and in particular stopping or reversing the progress of the disease. The desired response in the treatment of a disease or condition may also be a delay in the onset or prevention of the onset of the disease or said condition. In particular, the term "effective amount" refers to an amount of a therapeutic agent that is sufficient to prevent the development, recurrence or occurrence of cancer and one or more of its symptoms, reduce the severity, duration of cancer, reduce one or more symptoms of malignancy, prevent the development oncological disease, regression of oncological disease and/or prevention of metastasis of a malignant neoplasm. In an embodiment of the invention, the amount of therapy is effective to achieve stabilization, reduction, or elimination of the tumor stem cell population and/or eradication, removal, or control of primary cancer, metastatic cancer, and/or recurrent cancer.

The effective amount of an agent or composition described herein will depend on the condition being treated, the severity of the disease, individual patient parameters including age, physiology, size and weight, duration of treatment, type of concomitant therapy (if any), specific route of administration and similar factors. Accordingly, the administered doses of the agents described herein may depend on several such parameters. In the event that the response of the patient is insufficient when using the initial dose, higher doses (or effective higher doses achieved by another, more localized route of administration) can be used.

The agents and compositions provided herein may be used alone or in combination with conventional therapeutic regimens such as surgery, radiation, chemotherapy, and/or bone marrow transplantation (autologous, isogenic, allogeneic, or unrelated).

Cancer treatment is an area where combination strategies are particularly desirable because often the combined action of two, three, four or even more anticancer drugs/therapies creates synergistic effects that are significantly stronger than those of a monotherapeutic approach. Thus, in another embodiment of the present invention, the treatment of cancer can be effectively combined with various other drugs. These include, for example, combinations with conventional antitumor therapies, multiple epitope strategies, complementary immunotherapy, and treatment approaches targeting angiogenesis or apoptosis (for a review see, for example, Andersen et al. 2008: Cancer treatment: the combination of vaccination with other therapies Cancer Immunology Immunotherapy, 57(11): 1735-1743). Sequential administration of different agents may inhibit tumor cell growth at different checkpoints, while other agents may, for example, inhibit neoangiogenesis, cancer cell survival, or metastasis, potentially turning cancer into a chronic disease.

The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the invention.

Examples

Example 1 Descriptive Analysis of Genetic Immune Polymorphisms

The individual profile of single nucleotide polymorphisms (SNPs) in a patient's genome can be predictive of the rate of response to the therapeutic antibody IMAB362. To investigate these SNP patterns, all patients were genotyped for a range of SNPs with a known or suspected role in immune response and susceptibility to gastric cancer.

In particular, the following issues were considered:

- SNP genotypes of each patient in relation to the studied polymorphisms.

- The frequency of SNP genotypes in the patient population.

- Identification of patients with polymorphisms that can directly interfere with the action of IMAB362 (polymorphisms of the Fc receptor and the complement system).

- Accumulation of SNP genotypes per patient described as risk factors for susceptibility to gastric cancer, progression or treatment of malignancy.

- Correlation of SNP genotypes with clinical outcome.

- Correlation of SNP genotypes with progression-free survival (PFS).

All patients in cohorts 1, 2 and 3 were analyzed for genetic polymorphisms. Patient blood samples were collected on day 1 (V2a, preinfusion).

Whole blood samples (9 ml, EDTA-Monovette) were collected from all patients. EDTA blood was stored in 1 ml aliquots immediately after sampling at the study center at -20°C. EDTA blood samples were placed on dry ice (-70°C) and stored at -20°C. Upon arrival, blood samples were stored immediately at -20°C until DNA extraction.

SNPs of interest were selected through a literature search, focusing on SNPs known to affect the functioning of the immune system, and especially SNPs that have been described to influence the mechanism of action of therapeutic antibodies, such as Fc receptor and systemic polymorphisms. complement. Were selected and studied SNP, which are described to affect the survival of patients with gastric cancer, susceptibility to (gastric) malignancy or the progression of gastric cancer.

Genetic polymorphisms were analyzed with SNP Genotyping TaqMan™ assays (46 standard, 5 custom, Life Technologies) on the Fluidigm Biomark™ real-time PCR analysis platform. DNA isolation was performed according to standard protocols for isolation of genomic DNA from whole blood. The Fluidigm Biomark™ real-time PCR platform allows genotyping of up to 96 patient samples with 96 SNPs in a single measurement, as patient samples and specific SNP primers are applied to a laboratory chip with 96 channels for patient DNA samples and 96 orthogonal channels for SNP analyses. . The patient's genomic DNA was pre-amplified using Specific Target Amplification (STA). The pre-amplified DNA was subjected to TaqMan™ real-time PCR analysis under standard conditions on a Fluidigm Biomark™ real-time PCR platform. Allelic determination of SNPs was performed for each patient and each assay using proprietary Fluidigm software and "R" statistical analysis software. A subset of SNPs was confirmed by classical Sanger sequencing because Fluidigm results were ambiguous.

Genetic polymorphisms of 51 single nucleotide polymorphisms (SNPs) were identified in 53 patients. A blood sample from 1 patient did not allow sufficient DNA extraction for SNP analysis. 6 SNP genotypes were identified only for a subgroup of 20 patients. The genotype for the MDM2 SNP rs2279744 was not determined in 9 patients due to technical issues. The result of PTGS2 rs20417 genotyping in 1 patient was inconclusive and was not investigated further.

Determining the matrix of SNP genotypes for test patients allows statistical testing of the patient population for genotype frequency shifts compared to genotype frequency in European control populations. SNP genotype frequencies in European control populations are based on data collected by international SNP genotyping projects (HapMap-CEU, PGA-EUROPEAN-PANEL, CAUC1, pilot_1_CEU_low_coverage_panel, CEU_GENO_PANEL, PDR-90), which are deposited in the public database dbSNP (National Center Biotechnology Information, Bethesda, MD, USA The number of patients per genotype of a given SNP was compared with the number of patients per genotype in European control populations The number of patients per genotype of control populations was calculated by multiplying the reported relative frequency of the SNP genotype in the population by the reported number of samples tested. This allowed a direct chi-square fit test to identify statistically significant differences between the patient population and the corresponding control population.

A chi-square test was performed for 48 of the 51 SNPs examined. SNP genotype frequencies are not deposited in public databases, except for C1QA (rs1044378), FCGR2C (Q57X (C→T)), and MDM2 (rs2279744) SNPs. SNPs with a statistically significant genotype frequency shift between the patient and control populations (5 out of 48 SNPs, p<0.05) are shown in FIG. one.

4 of these 5 SNPs have been shown to play a role in susceptibility to gastric malignancy/cancer. All 4 malignancy/gastric cancer SNPs indeed show overrepresentation of the respective malignancy-associated genotype in the patient population, as expected for patients with gastric cancer (Table 1).

To date, 1 of these 5 SNPs (rs12146727 (C1S)) has not been shown to be a risk/susceptibility factor for malignancy or gastric cancer. This SNP has so far only been described once as a putative risk factor for cardiovascular disease.

Table 1. SNPs associated with susceptibility to gastric cancer with statistically significant differences in genotype frequency between patient and control populations

Gene SNP number overrepresented
genotype Major genotype at risk for malignant neoplasm (stomach) susceptibility CDH1 rs16260 AA AA IL2 rs2069762 GG GG PLCE1 rs2274223 GG GG CTLA4 rs231775 GG GG

Of the 51 SNPs studied in the patient population, 49 show a variant allelic pattern in the studied patient population. This allows SNP allelic frequency shifts to be tested between patient subpopulations, which ideally can help determine the intended responder population. Only 2 SNPs, C1QA (rs1044378) and FCGR2C (AHN1ME8) show an invariant SNP genotype in all patients, precluding any type of differential analysis. For 5 SNPs, a statistically significant shift in allelic frequency could be determined in this study compared to control populations, providing evidence for the principle that SNP allelic frequency depends on the composition of a given population. Thus, the selection of testable SNPs is well suited for the future identification of SNPs as biomarker candidates.

Polymorphisms of the Fc receptor and the complement system can directly influence the mode of action of IMAB362. Patients were genotyped for SNP alleles in genes that can influence the efficacy of antibody-based therapy, such as FCGR3A (F176V[T→G], rs396991), FCGR2A (H131R [T→C], rs1801274), and C1QA ([276A→ G], rs172378) (Table 2).

Patients were additionally genotyped for the published SNP alleles of the FCGR2C gene (Q57X [C→T], no rs number) and for the complement system factors C1S (R119H [G→A], rs12146727) and C1QA (rs292001, rs1044378). These SNPs have not previously been shown to affect antibody therapy, but they have been included as interesting candidate SNPs.

Table 2. Patients with polymorphisms of the Fc receptor and the complement system. The SNP genotypes of patients with well-known polymorphisms of the Fc receptor and the complement system are presented. FCGR3A Val/Val polymorphisms with a putative positive effect on antibody therapy are in bold and underlined. Polymorphisms in FCGR2A (Arg/Arg) and C1QA [G/G] with a presumed negative effect on antibody therapy are shown in gray and in bold.

A total of 23 patients exhibit at least one of the well-documented Fc receptor and complement polymorphisms. 4 patients (100411, 101120, 200336 and 400109) were homozygous for the FCGR3A (F176V [T→G]) allele, which is reported to increase response rate and progression-free survival with antibody therapy. 12 patients are homozygous for the FCGR2A allele (H131R [T→C]), another 10 patients are homozygous for the C1QA allele ([276A→G]). Both of these SNPs have been shown to have a negative effect on antibody therapy. A total of 21 patients are homozygous for both the FCGR2A allele (H1 31R [T→C]) and the C1QA allele ([276A→G] (patient 100511 is homozygous for both SNP alleles).

The correlation of the above data with the progression of patients' disease may provide insight into the role of Fc receptor polymorphisms and the complement system for treatment with IMAB362.

Disease progression and the efficacy of antibody treatment in patients may be affected by the accumulation of SNPs described as risk factors for gastric cancer predisposition, progression, or treatment of malignancies. Among the 51 SNPs examined, up to 43 SNPs allow classification of the corresponding SNP genotypes as “at risk” versus “without risk” genotypes. The number of homozygous SNP genotypes that are risk factors per patient was counted, since they are described in general as the most important risk alleles. The relative frequency of the number of homozygous genotypes at risk per patient relative to the number of risk factor SNPs studied per patient is shown in FIG. 2.

An accumulation of 14-46% of the studied risk genotypes per patient was observed. This broad distribution allows one to investigate whether the accumulation of risk SNP genotypes per patient correlates with the patient's clinical outcome.

Thus, 53 out of 54 patients were successfully genotyped for 51 SNPs. 49 out of 51 SNPs demonstrate a variant of the SNP allele, which allows analysis of patient subpopulations for a significant change in the frequency of the SNP genotype. Homozygous Fc receptor and complement system polymorphisms described as modulators of antibody therapy were found in 23 of 53 patients. An accumulation of 14 to 46% of the studied risk genotypes per patient was observed.

Example 2 Correlation of SNP Genotyping with Clinical Outcomes

The goal of correlating clinical outcomes with genetic polymorphism genotypes is to identify putative candidate SNP biomarkers predictive of patient clinical outcomes. The putative biomarker candidates identified in this analysis will be tested in subsequent phase IIb and phase III studies. Testing putative biomarker candidates in Phase IIb will allow differentiation between putative prognostic and predictive SNP candidates.

Correlation analyzes for each SNP with clinical outcome were performed independently for two defined patient populations in phase IIa clinical trials: a "complete analysis group" (FAS) population of 40 patients and a "per protocol group" population of 21 patients.

The absolute genotype frequencies of the corresponding SNP for each clinical outcome group ("responder", "nonresponder") of the patient population were quantified using SAS Enterprise Guide 6.1. Absolute genotype frequencies were organized in study population statistical structure tables (3×2 or 2×2) structured by clinical outcome and SNP genotype. Pearson's chi-square test was used as a standard statistical test. Fisher's exact test was used in some cases for 2×2 tables of the statistical structure of the study population, if the numerical structure of the dataset prohibited the use of Pearson's chi-square test. The level of statistical significance was p<0.05. Correlation analysis was performed using SAS Enterprise Guide 6.1 statistical analysis software.

To explore the effect of SNP genotypes on progression-free survival, Kaplan-Meier curves were calculated for each group and then formally compared using the statistical Logrank test. The level of statistical significance was p<0.05. Logrank statistics were implemented using SAS Enterprise Guide 6.1 statistical analysis software.

Correlation of clinical outcome with SNP genotyping was performed to determine putative prognostic or prognostic candidate SNP biomarkers. The correlation was studied in two groups of patients, the FAS population and the PP population.

The FAS population includes 40 patients, 12 patients defined as "responder" (clinical outcome "partial remission" or "stable disease"), and 28 patients as "non-responder" (clinical outcome "progression of the disease"). One patient sample (100801, non-responder) of the FAS population was not available for SNP analysis as described above, so the maximum number of FAS patients analyzed for correlations was reduced to 39. The PP population includes 21 patients with 10 responder patients and 11 patients non-responders.

The number of patients tested for SNP varies from 20 to 39 (in the FAS population) and from 20 to 21 (in the PP population).

Correlation analysis was performed as described above. Overall, of the 51 SNPs studied, 2 showed a statistically significant correlation with clinical outcome in the FAS population as well as in the PP population.

The 2 SNPs that demonstrated a statistical correlation between clinical outcome and the corresponding SNP genotype in both populations are FCGR2A rs1801274 (p = 0.0004 [PP], p = 0.008 [FAS]) and IL-10 rs1800896 (p = 0.042 [PP], p = 0.022 [FAS]) (Table 3). The number of patients statistically tested for SNPs was 21 (PP) and 39 (FAS) for each of these 2 SNPs.

Table 3. SNPs demonstrating a statistical correlation between clinical outcome and SNP genotype in the PP population as well as in the FAS population.

RS number Gene name Genotype overrepresented in the responder population p-value (PP) p-value (FAS) rs1801274 FCGR2A [CT] 0.0004 0.008 rs1800896 IL10 [GG] 0.042 0.022

(Chi-square test, statistically significant: p<0.05)

5 SNPs show a correlation with clinical outcome in the same patient population (FAS or PP), which can be demonstrated for DNMT3A rs1550117 [PP, p = 0.035], SMAD4 rs12456284 [FAS, p = 0.02], MUC1 rs4072037 (FAS, p = 0.03), EGF rs4444903 [FAS, p = 0.049], and CDH1 rs16260 [FAS p = 0.049]) (Table 4).

(Chi-square test, statistically significant: p<0.05)

Studying the overrepresentation or underrepresentation of SNP genotypes in responder/nonresponder patients can provide a scientific explanation for statistically significant differences in frequency.

Genotypes of two SNPs, rs11615 (ERCC1) and rs396991 (FCGR3A), correlate with prolonged progression-free survival (PFS) in the PP population (Table 5).

Table 5. SNPs showing statistical correlation between prolonged PFS and SNP genotypes in the PP population

RS number Gene name The genotype that correlates
with PFS P-value (PP) P-value (FAS) rs11615 ERCC1 [TT] 0.0001 0.13 rs396991 FCGR3A [TG]/[TT] 0.0007 0.25

The number of patients statistically tested for SNPs was 21 (PP) and 39 (FAS) for each of the 9 SNPs listed.

FCGR2A rs1801274 [C/T]: In PP, all patients carrying the heterozygous rs1801274 [CT] genotype are indeed responders (8), as reflected in the highly significant p-value (0.0004) of the statistical test. All patients with PR (4 out of 4) have this genotype. The majority of non-responders (73%, 8 out of 11) show the homozygous [TT] genotype (Table 6). This genotype distribution pattern can also be found in the FAS population, although not as scatter as in the PP population (Table 7). Quite a few non-responders in the FAS population also support the [CT] genotype (30%), resulting in a less pronounced, but still statistically highly significant, p-value.

Table 6 List of rs1801274 (FCGR2A) genotypes in PP patients and corresponding frequencies in responder (PR and SD) and non-responder (PD) patients

Genotype rs1801274 (FCGR2A) (Result) best answer PFS
[Days] Abs. frequency [ST] Rel. frequency [ST] 100702 CT RESP PR 322 eight 80% 200316 CT RESP PR 302 100603 CT RESP PR 287 200315 CT RESP PR 238 100108 CT RESP SD 330 100124 CC RESP SD 170 100709 CT RESP SD 146 101302 CT RESP SD 141 101109 TT RESP SD 132 100534 CT RESP SD 78 101116 TT NONRESP PD 114 0 0% 100510 TT NONRESP PD 112 200310 CC NONRESP PD 102 200319 CC NONRESP PD 73 101105 TT NONRESP PD 71 100411 TT NONRESP PD 70 100513 TT NONRESP PD 70 100605 CC NONRESP PD 70 400109 TT NONRESP PD 67 400101 TT NONRESP PD 65 101120 TT NONRESP PD 64

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Table 7. List of rs1801274 (FCGR2A) genotypes

Genotype rs1801274 (FCGR2A) (Result) best answer PFS
[Days] Abs. frequency [ST] Rel. frequency [ST] 100702 CT RESP PR 322 ten 83% 200316 CT RESP PR 302 100603 CT RESP PR 287 200315 CT RESP PR 238 200205 CT RESP SD 476 100108 CT RESP SD 330 400112 CT RESP SD 194 100124 CC RESP SD 170 100709 CT RESP SD 146 101302 CT RESP SD 141 101109 TT RESP SD 132 100534 CT RESP SD 78 100715 CC NONRESP PD 141 eight thirty% 100804 CT NONRESP PD 119 101116 TT NONRESP PD 114 100510 TT NONRESP PD 112 100808 TT NONRESP PD 112 200310 CC NONRESP PD 102 200336 CC NONRESP PD 90 101201 CT NONRESP PD 79 200207 TT NONRESP PD 75 200319 CC NONRESP PD 73 101105 TT NONRESP PD 71 100411 TT NONRESP PD 70 100513 TT NONRESP PD 70 100605 CC NONRESP PD 70 400109 TT NONRESP PD 67 400101 TT NONRESP PD 65 101120 TT NONRESP PD 64 400111 CT NONRESP PD 60 100901 CT NONRESP PD 55 100529 TT NONRESP PD fifty 100127 CC NONRESP PD 47 100410 CT NONRESP PD 46 100518 CT NONRESP PD 35 100310 CC NONRESP PD thirty 100607 CT NONRESP PD 27 100711 CC NONRESP PD 22 101007 CT NONRESP PD 17

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

FCGR2A rs1801274 [C/T] Survival Analysis: Significant overrepresentation of the rs1801274 [CT] genotype in the responder population was expected to be correlated with prolonged progression-free survival (PFS). Indeed, in both the PP (Fig. 3) and FAS (Fig. 4) populations, the [CT] genotype also significantly correlates with prolonged PFS (PP p = 0.0007, FAS p = 0.03). Interestingly, however, during the first 60 days of treatment, FAS patients with the [TT] genotype tend to have a higher degree of PFS than those with the [CC] or [CT] genotype. Thus, the survival analysis confirms that rs1801274 (FCGR2A) is a very interesting putative candidate biomarker of a predictive or prognostic nature.

IL-10 rs1800896 [A/G]: In PP, none of the non-responder patients have the homozygous rs1800896 [GG] genotype (Table 8). This genotype occurs at an increased frequency (40%) in responder patients (4 out of 10). Only 1 out of 10 responders (10%) show the [AA] genotype, the rest of the responders show the heterozygous [GA] genotype. In FAS, a comparable genotype frequency distribution can be observed (Table 9), although in this population, the [GG] genotype can be observed in non-responder patients at a low frequency (11%, 3 of 27).

Table 8 List of rs1800896 (IL-10) genotypes in PP patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Genotype rs1800896 (IL-10) (clinical outcome) best answer PFS
[Days] Abs. frequency [GG] Rel. frequency [GG] 100702 AA RESP PR 322 four 40% 200316 GG RESP PR 302 100603 GA RESP PR 287 200315 GA RESP PR 238 100108 GG RESP SD 330 100124 GA RESP SD 170 100709 GA RESP SD 146 101302 GA RESP SD 141 101109 GG RESP SD 132 100534 GG RESP SD 78 101116 AA NONRESP PD 114 0 0% 100510 AA NONRESP PD 112 200310 GA NONRESP PD 102 200319 GA NONRESP PD 73 101105 GA NONRESP PD 71 100411 AA NONRESP PD 70 100513 AA NONRESP PD 70 100605 GA NONRESP PD 70 400109 GA NONRESP PD 67 400101 AA NONRESP PD 65 101120 GA NONRESP PD 64

RESP: Responder, NONRESP: Nonresponder, PFS: Progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Table 9 List of rs1800896 (IL-10) genotypes in FAS patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Genotype rs1800896 (IL-10) Clinical outcome best answer PFS
[Days] Abs. frequency [GG] Rel. frequency [GG] 100702 AA RESP PR 322 6 fifty% 200316 GG RESP PR 302 100603 GA RESP PR 287 200315 GA RESP PR 238 200205 GG RESP SD 476 100108 GG RESP SD 330 400112 GG RESP SD 194 100124 GA RESP SD 170 100709 GA RESP SD 146 101302 GA RESP SD 141 101109 GG RESP SD 132 100534 GG RESP SD 78 100715 GA NONRESP PD 141 3 eleven% 100804 AA NONRESP PD 119 101116 AA NONRESP PD 114 100510 AA NONRESP PD 112 100808 GG NONRESP PD 112 200310 GA NONRESP PD 102 200336 AA NONRESP PD 90 101201 GA NONRESP PD 79 200207 GA NONRESP PD 75 200319 GA NONRESP PD 73 101105 GA NONRESP PD 71 100411 AA NONRESP PD 70 100513 AA NONRESP PD 70 100605 GA NONRESP PD 70 400109 GA NONRESP PD 67 400101 AA NONRESP PD 65 101120 GA NONRESP PD 64 400111 GA NONRESP PD 60 100901 GG NONRESP PD 55 100529 GA NONRESP PD fifty 100127 AA NONRESP PD 47 100410 GG NONRESP PD 46 100518 GA NONRESP PD 35 100310 GA NONRESP PD thirty 100607 GA NONRESP PD 27 100711 GA NONRESP PD 22 101007 GA NONRESP PD 17

RESP: Responder, NONRESP: Nonresponder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival Analysis of rs1800896 (IL-10) [A/G]: The rs1800896 [GG] genotype is significantly overrepresented in responder patients. Statistical correlation of the [GG] genotype with PFS shows that in the PP and FAS population, the [GG] genotype is not significantly correlated with PFS (PP p = 0.27 (Fig. 5), FAS p = 0.08 (Fig. 6)). However, the p-value for the FAS survival correlation borders on significance, which may indicate that in larger populations with reduced statistical noise, significance may well be reached. Overall, rs1800896 (IL-10) is an interesting putative biomarker candidate.

DNMT3A rs1550117 [G/A]: In PP, 4 responders (40%) show the [GA] genotype while all non-responders show the [GG] genotype (p = 0.03, Table 10).

Table 10 List of rs1550117 (DNMT3A) genotypes in PP patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Rs1550117 (DNMT3A) clinical outcome best answer PFS
[Days] Abs. frequency [GA] Rel. frequency [GA] 1007-02 GG RESP PR 322 four 40% 2003-16 GG RESP PR 302 1006-03 GA RESP PR 287 2003-15 GA RESP PR 238 1001-08 GG RESP SD 330 1001-24 GG RESP SD 170 1007-09 GA RESP SD 146 1013-02 GA RESP SD 141 1011-09 GG RESP SD 132 1005-34 GG RESP SD 78 1011-16 GG NONRESP PD 114 0 0% 1005-10 GG NONRESP PD 112 2003-10 GG NONRESP PD 102 2003-19 GG NONRESP PD 73 1011-05 GG NONRESP PD 71 1004-11 GG NONRESP PD 70 1005-13 GG NONRESP PD 70 1006-05 GG NONRESP PD 70 4001-09 GG NONRESP PD 67 4001-01 GG NONRESP PD 65 1011-20 GG NONRESP PD 64

RESP: Responder, NONRESP: Nonresponder, PFS: Progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival Analysis of rs1550117 (DNMT3A) [G/A]: The rs1550117 [GA] genotype is significantly overrepresented in responder patients in the PP population. In the FAS population, the difference in PFS between [GA] and [GG] carriers is of borderline significance (FAS p = 0.058) (Fig. 7).

In the FAS population, only one patient is a carrier of the [AA] genotype.

SMAD4 rs12456284 [G/A]: In FAS, a statistically significant overrepresentation of the [GA] genotype (7 of 12 patients, 58%) can be observed compared to the [AA] and [GG] genotypes in the responder population (p = 0.023, Table 11) . In a population of FAS non-responders, a [GA] genotype frequency of 19% (5 out of 27 non-responders) can be observed. In the PP population, this relationship is indicated by the significance of the trend (p = 0.081, data not shown).

Table 11. List of rs12456284 (SMAD4) genotypes

Genotype rs12456284 (SMAD4) clinical outcome best answer PFS
[Days] Abs. frequency [GA] Rel. frequency [GA] 1007-02 AA RESP PR 322 7 58% 2003-16 GA RESP PR 302 1006-03 GA RESP PR 287 2003-15 GA RESP PR 238 2002-05 GA RESP SD 476 1001-08 AA RESP SD 330 4001-12 AA RESP SD 194 1001-24 GA RESP SD 170 1007-09 GA RESP SD 146 1013-02 AA RESP SD 141 1011-09 AA RESP SD 132 1005-34 GA RESP SD 78 1007-15 GG NONRESP PD 141 5 19 % 1008-04 AA NONRESP PD 119 1011-16 GA NONRESP PD 114 1005-10 AA NONRESP PD 112 1008-08 AA NONRESP PD 112 2003-10 GA NONRESP PD 102 2003-36 AA NONRESP PD 90 1012-01 AA NONRESP PD 79 2002-07 AA NONRESP PD 75 2003-19 AA NONRESP PD 73 1011-05 AA NONRESP PD 71 1004-11 AA NONRESP PD 70 1005-13 AA NONRESP PD 70 1006-05 AA NONRESP PD 70 4001-09 AA NONRESP PD 67 4001-01 AA NONRESP PD 65 1011-20 AA NONRESP PD 64 4001-11 AA NONRESP PD 60 1009-01 GA NONRESP PD 55 1005-29 AA NONRESP PD fifty 1001-27 AA NONRESP PD 47 1004-10 AA NONRESP PD 46 1005-18 AA NONRESP PD 35 1003-10 AA NONRESP PD thirty 1006-07 AA NONRESP PD 27 1007-11 GA NONRESP PD 22 1010-07 GA NONRESP PD 17

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival analysis of rs12456284 (SMAD4) [G/A]: The rs12456284 [GA] genotype is significantly overrepresented in FAS responders and shows the same trend in PP responders. Statistical correlation of rs12456284 genotypes with PFS demonstrates that in the PP population, the [GA] genotype is significantly correlated with PFS (PP p = 0.048) using the Gehan-Brelow-Wilcoxon test (Fig. 8), while the significance using the log-rank test is p = 0.35. The Gehan-Brelow-Wilcoxon test gives more weight to PFS events at early time points than the log-rank test, and indeed the difference between [GA] and [AA] carriers is most pronounced during the first 100 days of this Phase IIa clinical trial. In the FAS population, there is no significant correlation of [GA] genotype with PFS (p = 0.20 (log-rank), p = 0.23 (Gehan-Brelow-Wilcoxon)), although visual inspection indicates a tendency for [GA] carriers to have prolonged PFS .

MUC1 rs4072037 [A/G]: In FAS, the rs4072037 genotype with the highest frequency of 67% in the responder population is [AA] (8 of 12), while among non-responders this genotype is expressed in only 26% of patients (7 of 27). Neither one of the responder patients does not show the homozygous [GG] genotype (Table 12), while the non-responders show the [GG] genotype at a rate of 22% (6 of 27). This differential distribution of genotypes in responder and non-responder patients is statistically significant (p = 0.03). A comparable genotype distribution profile is found in the PP population (data not shown), where the responder exhibits approximately the same relative frequency of the [AA] genotype of 70% (7 out of 10) as in the FAS population (significance trend p = 0.11).

Table 12 List of rs4072037 (MUC1) genotypes in FAS patients and corresponding frequencies in responder patients (PR and SD) and nonresponder patients (PD)

Patient ID Genotype rs4072037 (MUC1) clinical outcome best answer PFS
[Days] Abs. frequency [AA] Rel. frequency [AA] 100702 AA RESP PR 322 eight 67% 200316 AA RESP PR 302 100603 AA RESP PR 287 200315 AG RESP PR 238 200205 AA RESP SD 476 100108 AA RESP SD 330 400112 AG RESP SD 194 100124 AA RESP SD 170 100709 AG RESP SD 146 101302 AA RESP SD 141 101109 AG RESP SD 132 100534 AA RESP SD 78 100715 AG NONRESP PD 141 7 26% 100804 AG NONRESP PD 119 101116 AA NONRESP PD 114 100510 AA NONRESP PD 112 100808 AG NONRESP PD 112 200310 AG NONRESP PD 102 200336 GG NONRESP PD 90 101201 AG NONRESP PD 79 200207 AG NONRESP PD 75 200319 AA NONRESP PD 73 101105 GG NONRESP PD 71 100411 AA NONRESP PD 70 100513 GG NONRESP PD 70 100605 AG NONRESP PD 70 400109 GG NONRESP PD 67 400101 GG NONRESP PD 65 101120 AG NONRESP PD 64 400111 GG NONRESP PD 60 100901 AG NONRESP PD 55 100529 AA NONRESP PD fifty 100127 AG NONRESP PD 47 100410 AG NONRESP PD 46 100518 AA NONRESP PD 35 100310 AG NONRESP PD thirty 100607 AG NONRESP PD 27 100711 AG NONRESP PD 22 101007 AA NONRESP PD 17

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival analysis of rs4072037 (MUC1) [A/G]: Significant overrepresentation of the rs4072037 [AA] genotype in responder patients may indicate a correlation of this genotype with PFS. Statistical testing found that in the PP and FAS population, the [AA] genotype was significantly correlated with PFS (PP p = 0.001, (Fig. 9), FAS p = 0.02 (Fig. 10)). This survival analysis confirms rs4072037 (MUC1 ) as a very interesting putative predictive or predictive biomarker candidate.

EGF rs4444903 [G/A]: in FAS, the rs4444903 [AA] genotype is significantly overrepresented (p = 0.049) in the responder population (5 of 12; 42%) compared to the non-responder population (3 of 27, 11%) (Table 13) . In the PP population, this skewed distribution is not statistically significant (p = 0.32, data not shown).

Table 13 List of rs4444903 (EGF) genotypes in FAS patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Patient ID Genotype rs4444903 (EGF) (Result) clinical outcome PFS
[Days] Abs. frequency [AA] Rel. frequency [AA] 1007-02 GA RESP PR 322 5 42% 2003-16 AA RESP PR 302 1006-03 AA RESP PR 287 2003-15 AA RESP PR 238 2002-05 GA RESP SD 476 1001-08 GA RESP SD 330 4001-12 GA RESP SD 194 1001-24 AA RESP SD 170 1007-09 AA RESP SD 146 1013-02 GG RESP SD 141 1011-09 GA RESP SD 132 1005-34 GA RESP SD 78 1007-15 GG NONRESP PD 141 3 eleven% 1008-04 GA NONRESP PD 119 1011-16 AA NONRESP PD 114 1005-10 GA NONRESP PD 112 1008-08 GA NONRESP PD 112 2003-10 GA NONRESP PD 102 2003-36 GG NONRESP PD 90 1012-01 GG NONRESP PD 79 2002-07 GA NONRESP PD 75 2003-19 GG NONRESP PD 73 101105 AA NONRESP PD 71 1004-11 GA NONRESP PD 70 1005-13 GA NONRESP PD 70 1006-05 GA NONRESP PD 70 4001-09 GG NONRESP PD 67 4001-01 GG NONRESP PD 65 1011-20 GA NONRESP PD 64 4001-11 GA NONRESP PD 60 1009-01 GA NONRESP PD 55 1005-29 GG NONRESP PD fifty 1001-27 GG NONRESP PD 47 1004-10 GG NONRESP PD 46 1005-18 GG NONRESP PD 35 1003-10 GA NONRESP PD thirty 1006-07 GA NONRESP PD 27 1007-11 AA NONRESP PD 22 1010-07 GA NONRESP PD 17

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival analysis of rs4444903 (EGF) [G/A]: The correlation of the rs4444903 [AA] genotype with PFS in the PP or FAS population is not statistically significant (FAS p = 0.1, PP p = 0.16). However, a trend towards prolonged PFS is seen in both the PP and FAS populations (Fig. 11).

CDH1 rs16260 [C/A]: In FAS, the rs16260 [AA] genotype occurs at a significantly higher frequency in the responder population (5 of 12; 42%) than in the non-responder population (3 of 27, 11%) (P = 0.049 , table 14). In PP, this asymmetric distribution between both groups of patients is not significant (p = 0.72, data not shown).

Table 14. List of rs16260 (CDH1) genotypes in FAS patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Patient ID Genotype rs16260 (CDH1) clinical outcome best answer PFS
[Days] Abs. frequency [AA] Rel. frequency [AA] 1007-02 AA RESP PR 322 5 42% 2003-16 CC RESP PR 302 1006-03 CC RESP PR 287 2003-15 AA RESP PR 238 2002-05 AA RESP SD 476 1001-08 CA RESP SD 330 4001-12 AA RESP SD 194 1001-24 CC RESP SD 170 1007-09 AA RESP SD 146 1013-02 CC RESP SD 141 1011-09 CC RESP SD 132 1005-34 CC RESP SD 78 1007-15 CC NONRESP PD 141 3 eleven % 1008-04 CC NONRESP PD 119 1011-16 CA NONRESP PD 114 1005-10 AA NONRESP PD 112 1008-08 CC NONRESP PD 112 2003-10 CC NONRESP PD 102 2003-36 CA NONRESP PD 90 1012-01 CA NONRESP PD 79 2002-07 CC NONRESP PD 75 2003-19 CC NONRESP PD 73 1011-05 AA NONRESP PD 71 1004-11 AA NONRESP PD 70 1005-13 CC NONRESP PD 70 1006-05 CC NONRESP PD 70 4001-09 CA NONRESP PD 67 4001-01 CA NONRESP PD 65 1011-20 CC NONRESP PD 64 4001-11 CC NONRESP PD 60 1009-01 CC NONRESP PD 55 1005-29 CA NONRESP PD fifty 1001-27 CA NONRESP PD 47 1004-10 CA NONRESP PD 46 1005-18 CA NONRESP PD 35 1003-10 CC NONRESP PD thirty 1006-07 CA NONRESP PD 27 1007-11 CC NONRESP PD 22 1010-07 CC NONRESP PD 17

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival analysis of rs16260 (CDH1) [C/A]: The correlation of the rs16260 (CDH1) genotype with PFS bordered on statistical significance in the FAS population (log-rank p = 0.065, Gehan-Brelow-Wilcoxon test p = 0.032) (Fig. 12).

ERCC1 rs11615 [C/T]: PP showed a trend towards a higher frequency of the rs11615 [TT] genotype in the responder population (3 out of 10, 30%) (p = 0.068, non-responder population (0%)). Conversely, homozygous the [CC] genotype occurs only in the nonresponder population (2 patients) (Table 15).

Table 15 List of rs11615 (ERCC1) genotypes in PP patients and corresponding frequencies in responder patients (PR and SD) and non-responder patients (PD)

Patient ID Rs11615 (ERCC1) clinical outcome best answer PFS
[Days] Abs. frequency [TT] Rel. frequency [TT] 1007-02 CT RESP PR 322 3 thirty % 2003-16 CT RESP PR 302 1006-03 CT RESP PR 287 2003-15 TT RESP PR 238 1001-08 CT RESP SD 330 1001-24 TT RESP SD 170 1007-09 CT RESP SD 146 1013-02 CT RESP SD 141 1011-09 CT RESP SD 132 1005-34 TT RESP SD 78 1011-16 CT NONRESP PD 114 0 0% 1005-10 CT NONRESP PD 112 2003-10 CT NONRESP PD 102 2003-19 CT NONRESP PD 73 1011-05 CT NONRESP PD 71 1004-11 CT NONRESP PD 70 1005-13 CT NONRESP PD 70 1006-05 CT NONRESP PD 70 4001-09 CC NONRESP PD 67 4001-01 CT NONRESP PD 65 1011-20 CC NONRESP PD 64

RESP: responder, NONRESP: non-responder, PFS: progression-free survival, abs. frequency: absolute frequency, rel. frequency: relative frequency.

Survival Analysis of rs11615 (ERCC1) [C/T]: The rs11615 [TT] genotype was found exclusively in the responder population in the PP population. Statistical correlation of rs11615 genotypes with PFS demonstrates that the rs11615 genotype in PP correlates significantly with PFS, with [CT] and [TT] carriers showing prolonged survival compared to [CC] carriers (PP p = 0.0001) (Fig. 13). Despite this striking magnitude of significance, it should be noted that there were only 2 patients with the [CC] genotype and 3 patients with the [TT] genotype in PP. However, in the FAS population, the same effect can be observed as a trend (FAS p = 0.13, data not shown), suggesting that the effect is also valid for larger patient populations.

Survival analysis of FCGR3A rs396991 [T/G]: neither in PP nor in FAS did the rs396991 SNP genotype correlate with clinical outcome (FAS p = 0.49, PP p = 0.29, data not shown). However, a population survival analysis PP shows with high statistical significance that patients with [TG] and [TT] genotypes show improved PFS compared to [GG] (p = 0.0007, Fig. 14). This effect can also be observed in the FAS population (p = 0.25, data not shown). Despite the significance obtained for the PP population, it should be noted that only 3 PP patients are [GG] carriers.

Example 3 Discussion of Companion Immune Polymorphism Assays

The primary objective of this Phase IIa clinical trial was to evaluate the safety and efficacy of the anti-CLDN18.2 therapeutic monoclonal antibody IMAB362 in patients with gastroesophageal adenocarcinomas. In addition, concomitant genetic immune response polymorphism analyzes were performed to evaluate parameters that could serve as potential predictive or prognostic biomarkers in correlation with IMAB362 therapy.

Discussion of descriptive analysis of immune polymorphism

It has been shown that genetic polymorphisms in the patient's genome change the response rate of therapeutic antibodies. In order to investigate the effect of individual genetic variation on response rate, genotypes of 51 single nucleotide polymorphisms (SNPs) with a known or suspected role in immune response and susceptibility to or progression to gastric cancer were determined in patients.

In this study, 51 SNPs were successfully genotyped in 53 of 54 patients studied. A statistically significant shift in the frequency of genotypes in the patient population compared to the control populations can be found for 5 SNPs. Previously, 4 of these SNPs have been shown to be associated with predisposition to gastric malignancy/cancer. The respective genotypes of these SNPs associated with malignancy/gastric cancer are overrepresented in the study population, as would be expected in the GC-predominant patient population. An overrepresentation of the corresponding homozygous genotype may indicate a recessive mode of action suggesting impaired gene function as opposed to increased gene activity. This is supported by published data, for example, the gastric cancer-associated SNP rs16260 genotype in CDH1 is reported to downregulate CDH1 expression due to its position in the CDH1 promoter at -160.

Polymorphisms in genes involved in immune signaling have been investigated, even if these polymorphisms have not previously been described as risk factors for gastric cancer. Genetic polymorphisms in genes encoding immune signaling factors have been shown to significantly modulate the risk of developing gastric cancer. Thus, response rates to antibody-based therapy may be influenced by these SNPs.

An overrepresented IL-2 GG genotype (SNP rs2069762) in the patient population is associated with an increased risk of H. pylori infection-induced gastric atrophy and may predispose to gastric cancer. The CTLA4 SNP genotypes rs231775 and rs2274223 (PLCE1) have been described as risk factors for susceptibility to GC. Due to the fact that published studies on rs231775 are inconsistent on the genotype sequence, no conclusions will be drawn in this document.

In this study, polymorphisms of the Fcγ receptor and the complement system were investigated. A possible beneficial FCGR3A genotype encoding Val/Val [GG] was found in 4 APT patients, a FCGR2A genotype with potentially negative effects (Arg/Arg) [CC] could be found in 12 APT patients.

CDC, as a second effector mechanism, has been shown to be influenced by SNP polymorphisms: allelic carriers of the polymorphism in the complement component C1qA ([276A→G], rs172378) demonstrate a prolonged response after rituximab therapy for follicular lymphoma. A complement system polymorphism in C1QA with the 'GG' genotype was detected in 10 patients, possibly with a negative effect on response. However, the rs12146727 SNP polymorphism in the C1S complement component has so far only been described as being unrelated to antibody therapy or cancer.

The identification of significant genotype frequency shifts between patient and control populations demonstrates that SNP genotype frequency shifts can serve as prognostic and predictive markers in clinical trials.

The accumulation of risk SNP alleles may also affect the clinical outcome of patients. To enable this analysis, the number of homozygous genotypes with a risk SNP was counted per patient. The correlation of these amounts with response to therapy may provide insight into the role of risk factor SNP accumulation.

Discussion of the correlation of SNP genotyping with clinical outcome

FCGR2A rs1801274:

A study of over- or under-represented FCGR2A genotypes showed that in the PP population, all patients with the heterozygous rs1801274 [CT] genotype are responder patients and that partial response (PR) patients exclusively support this genotype. An overrepresented homozygous genotype in the non-responder-[TT] population. Simply observing these frequency distributions does not allow one to conclude whether the [CT] genotype is beneficial or whether the [TT] genotype is disadvantageous. In most studies examining the effects of SNP genotypes, the corresponding homozygous genotypes show the strongest biological effects, often indicating a recessive mode of action reflecting impaired gene function of both alleles as opposed to increased gene activity. In the event that SNP alleles lead to increased genetic activity, a stepwise effect of biological action can often be observed: one allele (i.e. heterozygous) increases gene activity, two alleles (i.e. homozygous) increase gene activity even more. In both cases, gain of function or loss of function, the strongest biological/clinical effects are usually observed in patients with homozygous genotypes. Under this assumption, overrepresentation of the homozygous [TT] genotype in the population of non-responders in the PP and FAS population may cause an adverse effect.

However, this is unexpected since the rs1801274 FCGR2A [TT] genotype has been described in a number of clinical studies as having a prolonging effect on PFS. In our Phase IIa clinical trial, a closer examination of the association between genotype and PFS in nonresponder patients revealed that FAS PD patients with the [TT] genotype show a real trend for longer PFS periods during the first 60 days of therapy, in contrast to FAS PD patients. with the [CT] genotype (compare Table 7 and Fig. 4). An interpretation to align this observation with [TT] underrepresentation in PFS-prolonged responders may be an overlap of two different molecular mechanisms: First, the rs1801274 [CT] genotype may be a marker for responder patients. This is a new observation that has not yet been described in the literature, and it can be assumed that this genotype is a prognostic marker for treatment with IMAB362. The molecular mechanism underlying this new observation is not yet clear.

The second observation, already described in the literature for other therapeutic antitumor antibodies, is the prolonged PFS of patients carrying the FCGR2A [TT] genotype. In our phase IIa study, this effect is due to an overlap of the postulated first mechanism observed only as a trend in non-responder patients. Mechanistically, the second observation can be explained by the increased binding affinity of the IgG1 antibody to its FCGR2A 131 His/His receptor allele (encoded by the [TT] genotype), in contrast to the weaker binding affinity to the homozygous ArgGRGA 131 Arg/Arg receptor allele (encoded by the [CC] genotype). ). In studies examining the effects of Fcγ receptor polymorphisms systematically, it has recently been shown that antibodies of the IgG1 isotype do indeed bind with different affinity to two allelic forms of the Fc2 IIA receptor, H131, with higher affinity than R131. It is generally believed that the differential affinity of IgG antibodies to FCGR2A receptor alleles influences the rate of triggering of effector mechanisms and hence prolonged PFS in patients carrying a high affinity receptor allele. Data supporting this hypothesis were provided by reports showing that the FCGR2A H131R and FCGR3A F176V (Phe>Val, rs396991) Fcγ receptor polymorphisms may influence the clinical efficacy of trastuzumab-based anti-IgG1 antibody therapy in patients with metastatic breast cancer. Patients with FCGR3A 176 Val/Val and FCGR2A 131 His/His genotypes showed significantly higher response rates and progression-free survival. The same polymorphisms have also been associated with the response rate of patients treated with rituximab (IgG1) for B-cell lymphoma. In another study, prolonged PFS after Cetuximab (IgG1) therapy may be associated with the FCGR3A 176 Val/Val genotype.

This is in contrast to recent well-established studies reporting no association between Fcγ receptor polymorphisms and survival, response rate, or progression-free survival for the antibodies in question. In the International Breast Cancer Research Group (BCIRG) study BCIRG-006, 1218 patients were treated in a randomized trial using two trastuzumab treatment groups and a non-trastuzumab control group. The associations described above between Fcγ receptor polymorphisms and trastuzumab efficacy cannot be confirmed. A long-term study with 460 patients using rituximab in combination with chemotherapy for follicular lymphoma found no association of Fcγ receptor polymorphisms with progression-free survival. In the REACH study with 419 patients, where patients received fludarabine and cyclophosphamide (FC) or rituximab plus FC, FCGR2A and FCGR3A polymorphisms had no significant effect on clinical outcome. Recent trials of cetuximab have also produced conflicting results, not recommending the use of Fcγ receptor polymorphisms as useful biomarkers. This may reflect differences in factors within a population or the effect of concurrent chemotherapy regimens.

MUC1 rs4072037:

MUC1 is a transmembrane glycoprotein of the mucin family. Mucins are high molecular weight proteins that are O-glycosylated in the N-terminal extracellular domain predominantly with oligosaccharides and n-glycan chains. Mucins are expressed on the apical surface of the epithelial lining of the respiratory and gastrointestinal tracts and ducts in the liver, pancreas, and kidneys. Transmembrane mucins span the membrane in a single α-helix and provide, through their sugar chains, a protective lining for the extracellular space. Mucins secreted into the extracellular space create a layer of mucous gel that serves as an additional physical protection for the epithelium.

Transmembrane MUC1 and secreted mucins MUC5C and MUC6 are the major mucins expressed in the stomach. MUC1 is translated as a single polypeptide chain that is self-cleaving. The N-terminal extracellular domain (MUC1-N) remains initially non-covalently associated with the transmembrane/intracytoplasmic domain (MUC1-C). This intracytoplasmic domain serves as a signaling domain that can enter the nucleus and associate with a number of transcription factors to directly activate gene expression. Cellular stress can lead to proteolytic cleavage of the MUC1-N and MUC1-C domain via a second proteolytic site. This can also be observed in cancer cells, where MUC1 is no longer expressed in an orderly manner on the apical membrane of the cell, but can be found overexpressed and localized throughout the cell. Cleavage of the extracellular domain (also known as CA15-3) in the extracellular space and intracellular localization of MUC1-C is a consequence. The intracytoplasmic signaling region acts as an oncogene, for example, by activating Wnt/β-catenin signaling and blocking apoptotic pathways.

However, the MUC1 extracellular domain is not only a static structural component, but also plays an important role during cell membrane signaling events. It has been demonstrated that the state of glycosylation and expression of the extracellular domain of MUC1 regulates the interaction of membrane signaling molecules and the extracellular matrix. Under-glycosylated MUC1-N in tumor cells has been reported to increase signaling between membrane molecules such as ICAM-1 or E-selectin and the nuclear protein MUC1. In addition, mucin expression and glycosylation state appear to mask membrane-associated molecules. In cancer cells, masking of HER2 proteins by mucin expression has been described as a possible mechanism for resistance to trastuzumab therapy.

The 'A' polymorphism of the MUC1 rs4072037 allele has been described as a risk factor for susceptibility to gastric cancer. This polymorphism is a G->A substitution in exon 2, resulting in alternative MUC1 splicing exactly at the predicted MUC1 signal peptide cleavage site. Deficient cleavage of the signal peptide can lead to aberrant MUC1 protein localization or an aberrant glycosylation pattern and consequently to deficient protein function.

In this phase IIa clinical trial, the rs4072037 [AA] genotype was found to be statistically associated with the responder population. It can be assumed that the insufficiently glycosylated or underexpressed [AA] allelic form of MUC1 provides better access for IMAB362 to the membrane target molecule CLDN18.2 expressed on cancer cells, which contributes to an increase in the effectiveness of treatment. This would make rs4072037 a predictive biomarker.

IL-10 rs1800896:

IL-10 is a key immune system regulator with pleiotropic functions. IL-10 is known to act as an anti-inflammatory, immunosuppressive cytokine, inhibiting macrophage-dependent antigen-specific T cell proliferation and macrophage-dependent cytokine production by T cells. However, IL-10 has also been described as an immunostimulatory cytokine that improves the differentiation and growth of B cells, granulocytes and mast cells, as well as the activation of NK cells and CD8 + T cells. The pleiotropic potential of IL-10 is also reflected in the broad expression of IL-10 in many types of immune cells, including Th2 cells, Treg cells, Th3 cells, NK cells, B cells, macrophages, and dendritic cells. This dual role of IL-10 is reflected in its tumor-promoting and tumor-inhibiting potential: IL-10, secreted by tumor cells or tumor infiltrating macrophage-type immune cells, allows tumor cells to elude immune surveillance through mechanisms that have only been elucidated. partially. One mechanism described involves Treg cells promoting the induction of peripheral tolerance through the expression of immunoregulatory cytokines such as IL-10. Another mechanism described is the inhibition of cross-presentation of tumor-specific antigens by dendritic cells and hence the prevention of T cells from initiating an effective immune response against tumor cells. On the other hand, exposure of malignant tumor cells to IL-10 leads to downregulation of HLA class I proteins, resulting in increased sensitivity to NK cell cytotoxicity.

The polymorphism of the IL-10 promoter rs1800896 at position (-1082) is of interest because the "G" allele is known to be a risk factor for gastric cancer and a risk factor for kidney cancer. The "G" allele of this polymorphism has been reported to be associated in vitro with decreased expression of IL-10 compared to the "A" allele. In responder patients of this Phase IIa clinical trial, the [GG] rs1800896 genotype is overrepresented, possibly indicating a lower level of relative expression of IL-10. It can be assumed that patients carrying the [GG] genotype have lower expression of IL-10, which, in turn, may make it more difficult for tumor cells to escape from immune surveillance using one of the mechanisms described above. Indeed, none of the 12 FAS responders showed elevated serum IL-10 levels, in contrast to 22% of FAS non-responders (6 of 27 examined). However, other authors state that the "A" allele is associated with a decrease in IL-10 expression.

It should also be noted that IL-10 signals through the intracellular mediator Stat3 and that Stat3 activation is MUC1-C dependent. Thus, the functional interaction of MUC1 and IL-10 may be why these molecules were statistically significant biomarker candidates in this phase IIa clinical trial. Finally, FCGR2A is expressed on macrophages, which are often the main source of IL-10 in the tumor microenvironment. If these putative biomarker candidates dominate current and future research, it may be of considerable interest to investigate the functional interaction of these factors.

Rs1550117 (DNMT3A):

Rs1550117 is a SNP in the DNMT3A gene encoding the enzyme DNA (cytosine-5)-methyltransferase 3A, which catalyses the transfer of methyl groups to specific CpG structures in DNA that induce epigenetic modification. The [AA] genotype has been shown to increase the risk of developing gastric cancer compared to [GG] or [GA]. In this study, [AA] was found in only one patient in the FAS population and in none of the patients in the PP population. This may indicate that [AA] is also a survival risk, preventing third and fourth treatments for [AA] carriers in this Phase IIa clinical trial. The finding that [GA] is significantly correlated with clinical outcome in PP suggests that this marker has potential as a predictive biomarker for IMAB362 treatment.

Rs12456284 (SMAD4):

Rs12456284 is an SNP in the SMAD4 gene encoding an intracellular TGFβ/BMP signaling co-transducer, 4 homologue of Mothers against decapentaplegic. It was published that the [GG] genotype significantly reduced the risk of developing stomach cancer. The statistically significant overrepresentation of the heterozygous [GA] genotype relative to the [AA] genotype in the FAS responder population suggests that this genotype may serve as a predictive biomarker. Prolonged PFS in patients in the PP population carrying [GA] is supporting evidence.

Rs4444903 (EGF):

It has been found that a functional rs4444903 polymorphism in the promoter region of the EGF gene modulates EGF protein levels, and that higher levels of EGF are detected in the serum of [GG] carriers. The G allele and [GG] genotype of this polymorphism showed significant correlations with an increased risk of developing gastrointestinal malignancies in a meta-analysis.

In this Phase IIa clinical study, the [AA] genotype is significantly overrepresented in FAS responders, and patients with this genotype show a trend towards prolonged PFS in the FAS and PP population. This may indicate that the rs4444903 [AA] genotype is a predictive or prognostic biomarker.

Rs16260 (CDH1):

The cell adhesion protein Cadherin (E-cadherin) is a member of the calcium dependent cadherin superfamily. Loss of function has been associated with cancer progression. The rs16260 [A] allele in the CDH1 promoter has been shown to reduce the transcriptional efficiency of cadherin 1. In addition, the -160A CDH1 allele has been described as a susceptibility factor to the development of gastric cancer.

In this study, carriers of the rs16260 [AA] genotype are statistically overrepresented in the FAS responder population. Based on this, it can be assumed that the [AA] genotype is a putative predictive biomarker.

Rs11615 (ERCC1) and rs396991 (FCGR3A):

The two SNPs rs11615 (ERCC1, "cross-complementing excision repair group 1" DNA repair protein) and rs396991 (FCGR3A, low-affinity immunoglobulin gamma Fc receptor type III-A) show genotype correlations (ERCC1 [TT], FCGR3A [TG] and [TT]) with prolonged PFS. Presumably, these SNPs are prognostic or predictive biomarkers.

link to applicant or agent file International Application No. 342-85

INSTRUCTIONS CONCERNING THE DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIALS

(PCT Rule 13 bis)

A. The indications below apply to the deposited microorganism or other biological material listed in the description on page 56, line 26.

B. DEPOSIT IDENTIFICATION Further deposits are indicated on the supplementary sheet

Name of depository institution

DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH

Address of depository institution (including postal code and country)

Mascheroder Weg 1b

38124 Braunschweig

DE

Deposit Date - Access Number

October 19, 2005 - DSM ACC2737

C. ADDITIONAL INSTRUCTIONS (leave blank if not applicable)

This information continues on an additional sheet.

- Mouse (Mus musculus) myeloma P3X63Ag8U. 1, fused with mouse (Mus musculus), splenocytes

Anti-human claudin-18A2 secreted by hybridoma

II DESIGNATED STATES FOR WHICH INSTRUCTIONS ARE GIVEN (unless the instructions apply to all the designated States)

E. SEPARATE INSTRUCTIONS SECTION (leave blank if not applicable)

The following indications will be submitted at a later date to the International Bureau (indicate the general nature of the indications, e.g. “deposit number”)

Receiving Office only - International Bureau only

This sheet was received with the international application - This sheet was received by the International Bureau

Authorized employee - Authorized employee

Form PCT/RO/134 (July 1998, reprinted January 2004)

New international patent application

Ganymed Pharmaceuticals AG, et al.

"Methods and compositions for predicting the therapeutic efficacy of the treatment of ONCOLOGICAL DISEASES and the prognosis of ONCOLOGICAL DISEASES"

Our reference: 342-85 PCT

_Supplementary sheet for biological material

Identification of further deposits:

1) Name and address of depository for deposits (DSM ACC2738, DSM ACC2739, DSM ACC2740, DSM ACC2741, DSM ACC2742, DSM ACC2743, DSM ACC2745, DSM ACC2746, DSM ACC2747, DSM ACC2748):

DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH

Mascheroder Weg 1b

38124 Braunschweig

DE

2) Name and address of depository for deposits (DSM ACC2808, DSM ACC2809, DSM ACC2810):

DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH

Inhoffenstr. 7B

38124 Braunschweig

DE

Deposit date Access numbers The directions below refer to the deposited microorganism in the description on the following pages(s) October 19, 2005 DSM ACC2738 Page 57, line 11 October 19, 2005 DSM ACC2739 Page 57, line 12 October 19, 2005 DSM ACC2740 Page 57, line 13 October 19, 2005 DSM ACC2741 Page 57, line 14 October 19, 2005 DSM ACC2742 Page 57, line 15 October 19, 2005 DSM ACC2743 Page 57, line 16 November 17, 2005 DSM ACC2745 Page 57, line 17 November 17, 2005 DSM ACC2746 Page 57, line 18 November 17, 2005 DSM ACC2747 Page 57, line 19 November 17, 2005 DSM ACC2748 Page 57, line 20 October 26, 2006 DSM ACC2808 Page 57, line 21 October 26, 2006 DSM ACC2809 Page 57, line 22 October 26, 2006 DSM ACC2810 Page 57, line 23

Additional guidance on all of the above deposits:

- Mouse (Mus musculus) myeloma P3X63Ag8U.1 fused with mouse (Mus musculus) splenocytes

-Hybridoma secreting anti-human claudin-18A2 antibody

3) Depositor:

All the above deposits were made:

Ganymed Pharmaceuticals AG

Freiligrathstrasse 12

55131 Mainz

DE

--->

SEQUENCE LIST

<110> Ganymed Pharmaceuticals AG

<120> METHODS AND COMPOSITIONS FOR PREDICTION OF THERAPEUTIC EFFICACY

OF CANCER TREATMENTS AND CANCER PROGNOSIS

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Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly

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Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg

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Gly Tyr Phe Thr Leu Leu Gly Leu Pro Ala Met Leu Gln Ala Val Arg

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Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val

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Ser Ile Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser

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Ala Lys Ala Asn Met Thr Leu Thr Ser Gly Ile Met Phe Ile Val Ser

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Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly Met Gly Gly

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Val Gly Trp Val Ala Gly Gly Leu Thr Leu Ile Gly Gly Val Met Met

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Cys Ile Ala Cys Arg Gly Leu Ala Pro Glu Glu Thr Asn Tyr Lys Ala

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Val Ser Tyr His Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly

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Leu Trp Arg Ser Cys Val Arg Gln Ser Ser Gly Phe Thr Glu Cys Arg

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Pro Tyr Phe Thr Ile Leu Gly Leu Pro Ala Met Leu Gln Ala Val Arg

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Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val

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Ser Ile Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser

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Ala Lys Ala Asn Met Thr Leu Thr Ser Gly Ile Met Phe Ile Val Ser

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Gly Leu Cys Ala Ile Ala Gly Val Ser Val Phe Ala Asn Met Leu Val

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Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly Met Gly Gly

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Val Gly Trp Val Ala Gly Gly Leu Thr Leu Ile Gly Gly Val Met Met

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Val Ser Tyr His Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly

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Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe

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Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly

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Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala

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Gly Phe Thr Glu Cys Arg Gly Tyr Phe Thr Leu Leu Gly Leu Pro Ala

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Met Leu Gln Ala Val Arg Ala

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Phe Ala Leu Lys Cys Ile Arg Ile Gly Ser Met Glu Asp Ser Ala Lys

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Ala Asn Met Thr Leu Thr Ser Gly

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Ala Asn Met Leu Val Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr

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Gly Ala Ala Leu Phe Val Gly Trp

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Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala

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Phe Gly Ala Ala Leu Phe Val Gly Trp

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Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

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Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

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Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

35 40 45

Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val

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100 105 110

Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

115 120 125

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

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Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

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165 170 175

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

180 185 190

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Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

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260 265 270

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

275 280 285

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

290 295 300

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

305 310 315 320

Ser Leu Ser Pro Gly Lys

325

<210> 14

<211> 466

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<213> Artificial

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Tyr Tyr Cys Ala Arg Tyr Asp Tyr Pro Trp Phe Ala Tyr Trp Gly Gln

115 120 125

Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val

130 135 140

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

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Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

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Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

180 185 190

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195 200 205

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

210 215 220

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

225 230 235 240

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

370 375 380

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly Lys

465

<210> 15

<211> 467

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Met Asp Trp Leu Trp Asn Leu Leu Phe Leu Met Ala Ala Ala Gln Ser

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Ile Gln Ala Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys

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Pro Gly Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe

35 40 45

Thr Asn Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu

50 55 60

Lys Trp Met Gly Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala

65 70 75 80

Glu Glu Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser

85 90 95

Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr

100 105 110

Tyr Phe Cys Ala Arg Leu Gly Phe Gly Asn Ala Met Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

130 135 140

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

145 150 155 160

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

165 170 175

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

180 185 190

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

195 200 205

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

210 215 220

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys

225 230 235 240

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

245 250 255

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

260 265 270

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

275 280 285

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

290 295 300

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

305 310 315 320

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

325 330 335

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

340 345 350

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

355 360 365

Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser

370 375 380

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

385 390 395 400

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

405 410 415

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

420 425 430

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

435 440 445

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

450 455 460

Pro Gly Lys

465

<210> 16

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Val His Ser Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg

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Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe

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Thr Asp Tyr Tyr Ile Asn Trp Val Lys Gln Arg Thr Gly Gln Gly Leu

50 55 60

Glu Trp Ile Gly Glu Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn

65 70 75 80

Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser

85 90 95

Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val

100 105 110

Tyr Phe Cys Ala Arg Ser Tyr Gly Ala Phe Asp Tyr Trp Gly Gln Gly

115 120 125

Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

130 135 140

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

145 150 155 160

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

165 170 175

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

180 185 190

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

195 200 205

Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro

210 215 220

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys

225 230 235 240

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

245 250 255

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

260 265 270

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

275 280 285

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

290 295 300

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

305 310 315 320

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

325 330 335

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

340 345 350

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

355 360 365

Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr

370 375 380

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

385 390 395 400

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

405 410 415

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

420 425 430

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

435 440 445

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

Lys

465

<210> 17

<211> 467

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<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

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Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly

1 5 10 15

Val His Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg

20 25 30

Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe

35 40 45

Thr Ser Tyr Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu

50 55 60

Glu Trp Ile Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn

65 70 75 80

Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser

85 90 95

Thr Ala Tyr Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val

100 105 110

Tyr Tyr Cys Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

130 135 140

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

145 150 155 160

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

165 170 175

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

180 185 190

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

195 200 205

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

210 215 220

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys

225 230 235 240

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

245 250 255

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

260 265 270

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

275 280 285

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

290 295 300

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

305 310 315 320

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

325 330 335

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

340 345 350

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

355 360 365

Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser

370 375 380

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

385 390 395 400

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

405 410 415

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

420 425 430

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

435 440 445

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

450 455 460

Pro Gly Lys

465

<210> 18

<211> 466

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 18

Met Glu Trp Arg Ile Phe Leu Phe Ile Leu Ser Gly Thr Ala Gly Val

1 5 10 15

His Ser Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro

20 25 30

Gly Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr

35 40 45

Asp Tyr Val Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu

50 55 60

Trp Ile Gly Glu Ile Tyr Pro Gly Ser Gly Ser Thr Tyr Tyr Asn Glu

65 70 75 80

Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr

85 90 95

Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr

100 105 110

Phe Cys Ala Arg Gly Val Leu Leu Arg Ala Met Asp Tyr Trp Gly Gln

115 120 125

Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

130 135 140

Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala

145 150 155 160

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

165 170 175

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

180 185 190

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

195 200 205

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

210 215 220

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

225 230 235 240

Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu

370 375 380

Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly Lys

465

<210> 19

<211> 469

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 19

Met Asp Trp Ile Trp Ile Met Leu His Leu Leu Ala Ala Ala Thr Gly

1 5 10 15

Ile Gln Ser Gln Val His Leu Gln Gln Ser Gly Ser Glu Leu Arg Ser

20 25 30

Pro Gly Ser Ser Val Lys Leu Ser Cys Lys Asp Phe Asp Ser Glu Val

35 40 45

Phe Pro Phe Ala Tyr Met Ser Trp Ile Arg Gln Lys Pro Gly His Gly

50 55 60

Phe Glu Trp Ile Gly Asp Ile Leu Pro Ser Ile Gly Arg Thr Ile Tyr

65 70 75 80

Gly Glu Lys Phe Glu Asp Lys Ala Thr Leu Asp Ala Asp Thr Val Ser

85 90 95

Asn Thr Ala Tyr Leu Glu Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala

100 105 110

Ile Tyr Tyr Cys Ala Arg Gly Glu Gly Tyr Gly Ala Trp Phe Ala Tyr

115 120 125

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly

130 135 140

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

145 150 155 160

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

165 170 175

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

180 185 190

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

195 200 205

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

210 215 220

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys

225 230 235 240

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

245 250 255

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

260 265 270

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

275 280 285

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

290 295 300

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

305 310 315 320

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

325 330 335

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

340 345 350

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

355 360 365

Gln Val Tyr Thr Leu Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln

370 375 380

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

385 390 395 400

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

405 410 415

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

420 425 430

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

435 440 445

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

450 455 460

Leu Ser Pro Gly Lys

465

<210> 20

<211> 240

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 20

Met Glu Ser Gln Thr Gln Val Leu Met Ser Leu Leu Phe Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr

20 25 30

Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr

100 105 110

Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr

115 120 125

Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

130 135 140

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

165 170 175

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

210 215 220

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235 240

<210> 21

<211> 235

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 21

Met His Phe Gln Val Gln Ile Phe Ser Phe Leu Leu Ile Ser Ala Ser

1 5 10 15

Val Ile Met Ser Arg Gly Gln Ile Val Leu Thr Gln Ser Pro Ala Ile

20 25 30

Met Ser Ala Ser Pro Gly Glu Lys Val Thr Ile Thr Cys Ser Ala Ser

35 40 45

Ser Ser Val Ser Tyr Met His Trp Phe Gln Gln Lys Pro Gly Thr Ser

50 55 60

Pro Lys Leu Trp Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro

65 70 75 80

Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile

85 90 95

Ser Arg Met Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg

100 105 110

Ser Ser Tyr Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

115 120 125

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

130 135 140

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

145 150 155 160

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

165 170 175

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

180 185 190

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

195 200 205

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

210 215 220

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 22

<211> 234

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 22

Met Glu Phe Gln Thr Gln Val Phe Val Phe Val Leu Leu Trp Leu Ser

1 5 10 15

Gly Val Asp Gly Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser

20 25 30

Thr Ser Val Gly Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn

35 40 45

Val Arg Thr Ala Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro

50 55 60

Lys Ala Leu Ile Tyr Leu Ala Ser Asn Arg His Thr Gly Val Pro Asp

65 70 75 80

Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

85 90 95

Asn Val Gln Ser Glu Asp Leu Ala Asp Tyr Phe Cys Leu Gln His Trp

100 105 110

Asn Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

115 120 125

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

130 135 140

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

145 150 155 160

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

165 170 175

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

180 185 190

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

195 200 205

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

210 215 220

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 23

<211> 240

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 23

Met Asp Ser Gln Ala Gln Val Leu Met Leu Leu Leu Leu Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala

20 25 30

Val Ser Val Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr

100 105 110

Tyr Cys Gln Gln Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr

115 120 125

Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

130 135 140

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

165 170 175

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

210 215 220

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235 240

<210> 24

<211> 240

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 24

Met Glu Ser Gln Thr Gln Val Leu Met Ser Leu Leu Phe Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr

20 25 30

Val Thr Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr

100 105 110

Tyr Cys Gln Asn Asp Tyr Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr

115 120 125

Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

130 135 140

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

165 170 175

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

210 215 220

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235 240

<210> 25

<211> 239

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 25

Met Asp Ser Gln Ala Gln Val Leu Ile Leu Leu Leu Leu Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala

20 25 30

Val Ser Ala Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Asn Ser Arg Thr Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr

100 105 110

Tyr Cys Lys Gln Ser Tyr Asn Leu Tyr Thr Phe Gly Gly Gly Thr Lys

115 120 125

Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro

130 135 140

Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu

145 150 155 160

Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp

165 170 175

Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp

180 185 190

Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys

195 200 205

Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln

210 215 220

Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235

<210> 26

<211> 240

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 26

Met Asp Ser Gln Ala Gln Val Leu Met Leu Leu Leu Leu Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala

20 25 30

Val Ser Val Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Ala Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr

100 105 110

His Cys Gly Glyn Gly Tyr Ser Tyr Pro Tyr Thr Phe Gly Gly Gly Thr

115 120 125

Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

130 135 140

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

165 170 175

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

210 215 220

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235 240

<210> 27

<211> 240

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 27

Met Asp Ser Gln Ala Gln Val Leu Met Leu Leu Leu Leu Trp Val Ser

1 5 10 15

Gly Thr Cys Gly Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala

20 25 30

Val Ser Val Gly Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser

35 40 45

Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr

100 105 110

Tyr Cys Gln Gln Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr

115 120 125

Lys Leu Glu Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe

130 135 140

Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys

145 150 155 160

Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val

165 170 175

Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln

180 185 190

Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser

195 200 205

Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His

210 215 220

Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230 235 240

<210> 28

<211> 234

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: chimeric monoclonal antibody

<400> 28

Met Glu Ser Gln Thr Leu Val Phe Ile Ser Ile Leu Leu Trp Leu Tyr

1 5 10 15

Gly Ala Asp Gly Asn Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser

20 25 30

Met Ser Val Gly Glu Arg Val Thr Leu Thr Cys Lys Ala Ser Glu Asn

35 40 45

Val Val Thr Tyr Val Ser Trp Tyr Gln Gln Lys Pro Glu Gln Ser Pro

50 55 60

Lys Leu Leu Ile Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp

65 70 75 80

Arg Phe Thr Gly Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser

85 90 95

Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr

100 105 110

Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg

115 120 125

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

130 135 140

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

145 150 155 160

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

165 170 175

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

180 185 190

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

195 200 205

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

210 215 220

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

225 230

<210> 29

<211> 117

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 29

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Tyr

20 25 30

Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Leu Pro Gly Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Asp Tyr Pro Trp Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala

115

<210> 30

<211> 118

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 30

Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Leu Gly Phe Gly Asn Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser

115

<210> 31

<211> 116

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 31

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Ser Tyr Gly Ala Phe Asp Tyr Trp Gly Gln Gly Thr Thr Leu

100 105 110

Thr Val Ser Ser

115

<210> 32

<211> 118

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 32

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe

50 55 60

Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Leu Thr Val Ser Ser

115

<210> 33

<211> 118

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 33

Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Val Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Tyr Pro Gly Ser Gly Ser Thr Tyr Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Gly Val Leu Leu Arg Ala Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser

115

<210> 34

<211> 120

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 34

Gln Val His Leu Gln Gln Ser Gly Ser Glu Leu Arg Ser Pro Gly Ser

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Asp Phe Asp Ser Glu Val Phe Pro Phe

20 25 30

Ala Tyr Met Ser Trp Ile Arg Gln Lys Pro Gly His Gly Phe Glu Trp

35 40 45

Ile Gly Asp Ile Leu Pro Ser Ile Gly Arg Thr Ile Tyr Gly Glu Lys

50 55 60

Phe Glu Asp Lys Ala Thr Leu Asp Ala Asp Thr Val Ser Asn Thr Ala

65 70 75 80

Tyr Leu Glu Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Ile Tyr Tyr

85 90 95

Cys Ala Arg Gly Glu Gly Tyr Gly Ala Trp Phe Ala Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ala

115 120

<210> 35

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 35

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser

20 25 30

Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu

100 105 110

Lys

<210> 36

<211> 106

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 36

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Phe Gln Gln Lys Pro Gly Thr Ser Pro Lys Leu Trp Ile Tyr

35 40 45

Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Ser Tyr Pro Pro Thr

85 90 95

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 37

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 37

Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly

1 5 10 15

Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile

35 40 45

Tyr Leu Ala Ser Asn Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser

65 70 75 80

Glu Asp Leu Ala Asp Tyr Phe Cys Leu Gln His Trp Asn Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 38

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 38

Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu

100 105 110

Lys

<210> 39

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 39

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser

20 25 30

Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile

100 105 110

Lys

<210> 40

<211> 112

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 40

Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala Val Ser Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser

20 25 30

Arg Thr Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asn Leu Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 41

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 41

Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr His Cys Gly Gln

85 90 95

Gly Tyr Ser Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

100 105 110

Lys

<210> 42

<211> 113

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 42

Asp Ile Val Met Ser Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu

100 105 110

Lys

<210> 43

<211> 107

<212> PRT

<213> Artificial

<220>

<223> Description of artificial sequence: Translation of PCR product

<400> 43

Asn Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly

1 5 10 15

Glu Arg Val Thr Leu Thr Cys Lys Ala Ser Glu Asn Val Val Thr Tyr

20 25 30

Val Ser Trp Tyr Gln Gln Lys Pro Glu Gln Ser Pro Lys Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly

50 55 60

Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Lys Ala

65 70 75 80

Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln Tyr Tyr Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210> 44

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 44

Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr

1 5 10 15

<210> 45

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 45

Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn

1 5 10 15

<210> 46

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 46

Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu

1 5 10 15

<210> 47

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 47

Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys

1 5 10 15

<210> 48

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 48

Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser

1 5 10 15

<210> 49

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 49

Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr

1 5 10 15

<210> 50

<211> 15

<212> PRT

<213> Artificial

<220>

<223> Epitope

<400> 50

Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly

1 5 10 15

<210> 51

<211> 467

<212> PRT

<213> Artificial Sequence

<220>

<223> chimeric monoclonal antibody

<400> 51

Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly

1 5 10 15

Val His Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg

20 25 30

Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe

35 40 45

Thr Ser Tyr Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu

50 55 60

Glu Trp Ile Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn

65 70 75 80

Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser

85 90 95

Thr Ala Tyr Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val

100 105 110

Tyr Tyr Cys Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

130 135 140

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

145 150 155 160

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

165 170 175

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

180 185 190

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

195 200 205

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

210 215 220

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

225 230 235 240

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

245 250 255

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

260 265 270

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

275 280 285

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

290 295 300

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

305 310 315 320

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

325 330 335

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

340 345 350

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

355 360 365

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

370 375 380

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

385 390 395 400

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

405 410 415

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

420 425 430

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

435 440 445

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

450 455 460

Pro Gly Lys

465

<210> 52

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be C or T

<400> 52

tgggatggag aaggtgggat ccaaanggga gaatttctgg gattttccat t 51

<210> 53

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or G

<400> 53

cccctaaacc cgcaacagtt gttacnggtt ctggtcatgc aagctctacc c 51

<210> 54

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or G

<400> 54

caacactact aaggcttctt tgggangggg aagtagggat aggtaagagg a 51

<210> 55

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or G

<400> 55

aattccacca gcacagccac tcactntgtg ctcatctcac tcctccagca g 51

<210> 56

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or G

<400> 56

aggtccagag ccagtgttct tgttcnacct gaaagtaatg gctctgggtt g 51

<210> 57

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or G

<400> 57

ctttcagccc caatccaagg gttgtngctg gaactttcca tcagttcttc c 51

<210> 58

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be A or C

<400> 58

ctagcaactc caggctagag ggtcancgcg tctatgcgag gccgggtggg c 51

<210> 59

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be C or T

<400> 59

atcccgtact gaagttcgtg cgcaangtgc cctgggaatt tggcgacgta a 51

<210> 60

<211> 51

<212> DNA

<213> Homo sapiens

<220>

<221> misc_feature

<222> (26)..(26)

<223> n may be G or T

<400> 60

cggctcctac ttctgcaggg ggcttnttgg gagtaaaaat gtgtcttcag a 51

<210> 61

<211> 2429

<212> DNA

<213> Homo sapiens

<400> 61

ctcttttcta agcttgtctc ttaaaaccca ctggacgttg gcacagtgct gggatgacta 60

tggagaccca aatgtctcag aatgtatgtc ccagaaacct gtggctgctt caaccattga 120

cagttttgct gctgctggct tctgcagaca gtcaagctgc agctccccca aaggctgtgc 180

tgaaacttga gcccccgtgg atcaacgtgc tccaggagga ctctgtgact ctgacatgcc 240

agggggctcg cagccctgag agcgactcca ttcagtggtt ccacaatggg aatctcattc 300

ccacccacac gcagcccagc tacaggttca aggccaacaa caatgacagc ggggagtaca 360

cgtgccagac tggccagacc agcctcagcg accctgtgca tctgactgtg ctttccgaat 420

ggctggtgct ccagacccct cacctggagt tccaggaggg agaaaccatc atgctgaggt 480

gccacagctg gaaggacaag cctctggtca aggtcacatt cttccagaat ggaaaatccc 540

agaaattctc ccatttggat cccaccttct ccatcccaca agcaaaccac agtcacagtg 600

gtgattacca ctgcacagga aacataggct acacgctgtt ctcatccaag cctgtgacca 660

tcactgtcca agtgcccagc atgggcagct cttcaccaat ggggatcatt gtggctgtgg 720

tcattgcgac tgctgtagca gccattgttg ctgctgtagt ggccttgatc tactgcagga 780

aaaagcggat ttcagccaat tccactgatc ctgtgaaggc tgcccaattt gagccacctg 840

gacgtcaaat gattgccatc agaaagagac aacttgaaga aaccaacaat gactatgaaa 900

cagctgacgg cggctacatg actctgaacc ccagggcacc tactgacgat gataaaaaaca 960

tctacctgac tcttcctccc aacgaccatg tcaacagtaa taactaaaga gtaacgttat 1020

gccatgtggt catactctca gcttgctgag tggatgacaa aaagagggga attgttaaag 1080

gaaaatttaa atggagactg gaaaaatcct gagcaaacaa aaccacctgg cccttagaaa 1140

tagctttaac tttgcttaaa ctacaaacac aagcaaaact tcacggggtc atactacata 1200

caagcataag caaaacttaa cttggatcat ttctggtaaa tgcttatgtt agaaataaga 1260

caaccccagc caatcacaag cagcctacta acatataatt aggtgactag ggactttcta 1320

agaagatacc tacccccaaa aaacaattat gtaattgaaa accaaccgat tgcctttatt 1380

ttgcttccac attttcccaa taaatacttg cctgtgacat tttgccactg gaacactaaa 1440

cttcatgaat tgcgcctcag atttttcctt taacatcttt tttttttttg acagagtctc 1500

aatctgttac ccaggctgga gtgcagtggt gctatcttgg ctcactgcaa acccgcctcc 1560

caggtttaag cgattctcat gcctcagcct cccagtagct gggattagag gcatgtgcca 1620

tcatacccag ctaatttttg tattttttat tttttttttt tagtagagac agggtttcgc 1680

aatgttggcc aggccgatct cgaacttctg gcctctagcg atctgcccgc ctcggcctcc 1740

caaagtgctg ggatgaccag catcagcccc aatgtccagc ctctttaaca tcttctttcc 1800

tatgccctct ctgtggatcc ctactgctgg tttctgcctt ctccatgctg agaacaaaat 1860

cacctattca ctgcttatgc agtcggaagc tccagaagaa caaagagccc aattaccaga 1920

accacattaa gtctccattg ttttgccttg ggatttgaga agagaattag agaggtgagg 1980

atctggtatt tcctggacta aattcccctt ggggaagacg aagggatgct gcagttccaa 2040

aagagaagga ctcttccaga gtcatctacc tgagtcccaa agctccctgt cctgaaagcc 2100

acagacaata tggtcccaaa tgactgactg caccttctgt gcctcagccg ttcttgacat 2160

caagaatctt ctgttccaca tccacacagc caatacaatt agtcaaacca ctgttattaa 2220

2280

ctatatgact tcagaaatgt taaaatagac taacctctaa caacaaatta aaagtgattg 2340

tttcaaggtg atgcaattat tgatgaccta ttttattttt ctataatgat catatattac 2400

ctttgtaata aaacattata accaaaaca 2429

<210> 62

<211> 317

<212> PRT

<213> Homo sapiens

<400> 62

Met Thr Met Glu Thr Gln Met Ser Gln Asn Val Cys Pro Arg Asn Leu

1 5 10 15

Trp Leu Leu Gln Pro Leu Thr Val Leu Leu Leu Leu Ala Ser Ala Asp

20 25 30

Ser Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro

35 40 45

Trp Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly

50 55 60

Ala Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn

65 70 75 80

Leu Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn

85 90 95

Asn Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser

100 105 110

Asp Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr

115 120 125

Pro His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His

130 135 140

Ser Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly

145 150 155 160

Lys Ser Gln Lys Phe Ser His Leu Asp Pro Thr Phe Ser Ile Pro Gln

165 170 175

Ala Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly

180 185 190

Tyr Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro

195 200 205

Ser Met Gly Ser Ser Ser Pro Met Gly Ile Ile Val Ala Val Val Ile

210 215 220

Ala Thr Ala Val Ala Ala Ile Val Ala Ala Val Val Ala Leu Ile Tyr

225 230 235 240

Cys Arg Lys Lys Arg Ile Ser Ala Asn Ser Thr Asp Pro Val Lys Ala

245 250 255

Ala Gln Phe Glu Pro Pro Gly Arg Gln Met Ile Ala Ile Arg Lys Arg

260 265 270

Gln Leu Glu Glu Thr Asn Asn Asp Tyr Glu Thr Ala Asp Gly Gly Tyr

275 280 285

Met Thr Leu Asn Pro Arg Ala Pro Thr Asp Asp Asp Lys Asn Ile Tyr

290 295 300

Leu Thr Leu Pro Pro Asn Asp His Val Asn Ser Asn Asn

305 310 315

<210> 63

<211> 1193

<212> DNA

<213> Homo sapiens

<400> 63

cgctccacct ctcaagcagc cagcgcctgc ctgaatctgt tctgccccct ccccacccat 60

ttcaccacca ccatgacacc gggcacccag tctcctttct tcctgctgct gctcctcaca 120

gtgcttacag ctaccacagc ccctaaaccc gcaacagttg ttacgggttc tggtcatgca 180

agctctaccc caggtggaga aaaggagact tcggctaccc agagaagttc agtgcccagc 240

tctactgaga agaatgcttt taattcctct ctggaagatc ccagcaccga ctactaccaa 300

gagctgcaga gagacatttc tgaaatgttt ttgcagattt ataaacaagg gggttttctg 360

ggcctctcca atattaagtt caggccagga tctgtggtgg tacaattgac tctggccttc 420

cgagaaggta ccatcaatgt ccacgacgtg gagacacagt tcaatcagta taaaacggaa 480

gcagcctctc gatataacct gacgatctca gacgtcagcg tgagtgatgt gccatttcct 540

ttctctgccc agtctggggc tggggtgcca ggctggggca tcgcgctgct ggtgctggtc 600

tgtgttctgg ttgcgctggc cattgtctat ctcattgcct tggctgtctg tcagtgccgc 660

cgaaagaact acgggcagct ggacatcttt ccagcccggg atacctacca tcctatgagc 720

gagtacccca cctaccacac ccatgggcgc tatgtgcccc ctagcagtac cgatcgtagc 780

ccctatgaga aggtttctgc aggtaatggt ggcagcagcc tctcttacac aaacccagca 840

gtggcagcca cttctgccaa cttgtaggggg cacgtcgccc gctgagctga gtggccagcc 900

agtgccattc cactccactc aggttcttca gggccagagc ccctgcaccc tgtttgggct 960

ggtgagctgg gagttcaggt gggctgctca cagcctcctt cagaggcccc accaatttct 1020

cggacacttc tcagtgtgtg gaagctcatg tgggcccctg agggctcatg cctgggaagt 1080

gttgtggtgg gggctcccag gaggactggc ccagagagcc ctgagatagc ggggatcctg 1140

aactggactg aataaaacgt ggtctcccac tgcgccaaaa aaaaaaaaaa aaa 1193

<210> 64

<211> 264

<212> PRT

<213> Homo sapiens

<400> 64

Met Thr Pro Gly Thr Gln Ser Pro Phe Phe Leu Leu Leu Leu Leu Thr

1 5 10 15

Val Leu Thr Ala Thr Thr Ala Pro Lys Pro Ala Thr Val Val Thr Gly

20 25 30

Ser Gly His Ala Ser Ser Thr Pro Gly Gly Glu Lys Glu Thr Ser Ala

35 40 45

Thr Gln Arg Ser Ser Val Pro Ser Ser Thr Glu Lys Asn Ala Phe Asn

50 55 60

Ser Ser Leu Glu Asp Pro Ser Thr Asp Tyr Tyr Gln Glu Leu Gln Arg

65 70 75 80

Asp Ile Ser Glu Met Phe Leu Gln Ile Tyr Lys Gln Gly Gly Phe Leu

85 90 95

Gly Leu Ser Asn Ile Lys Phe Arg Pro Gly Ser Val Val Val Gln Leu

100 105 110

Thr Leu Ala Phe Arg Glu Gly Thr Ile Asn Val His Asp Val Glu Thr

115 120 125

Gln Phe Asn Gln Tyr Lys Thr Glu Ala Ala Ser Arg Tyr Asn Leu Thr

130 135 140

Ile Ser Asp Val Ser Val Ser Asp Val Pro Phe Pro Phe Ser Ala Gln

145 150 155 160

Ser Gly Ala Gly Val Pro Gly Trp Gly Ile Ala Leu Leu Val Leu Val

165 170 175

Cys Val Leu Val Ala Leu Ala Ile Val Tyr Leu Ile Ala Leu Ala Val

180 185 190

Cys Gln Cys Arg Arg Lys Asn Tyr Gly Gln Leu Asp Ile Phe Pro Ala

195 200 205

Arg Asp Thr Tyr His Pro Met Ser Glu Tyr Pro Thr Tyr His Thr His

210 215 220

Gly Arg Tyr Val Pro Ser Ser Thr Asp Arg Ser Pro Tyr Glu Lys

225 230 235 240

Val Ser Ala Gly Asn Gly Gly Ser Ser Leu Ser Tyr Thr Asn Pro Ala

245 250 255

Val Ala Ala Thr Ser Ala Asn Leu

260

<210> 65

<211> 1629

<212> DNA

<213> Homo sapiens

<400> 65

acacatcagg ggcttgctct tgcaaaacca aaccacaaga cagacttgca aaagaaggca 60

tgcacagctc agcactgctc tgttgcctgg tcctcctgac tggggtgagg gccagcccag 120

gccagggcac ccagtctgag aacagctgca cccacttccc aggcaacctg cctaacatgc 180

ttcgagatct ccgagatgcc ttcagcagag tgaagacttt ctttcaaatg aaggatcagc 240

tggacaactt gttgttaaag gagtccttgc tggaggactt taagggttac ctgggttgcc 300

aagccttgtc tgagatgatc cagttttacc tggaggaggt gatgccccaa gctgagaacc 360

aagacccaga catcaaggcg catgtgaact ccctggggga gaacctgaag accctcaggc 420

tgaggctacg gcgctgtcat cgatttcttc cctgtgaaaa caagagcaag gccgtggagc 480

aggtgaagaa tgcctttaat aagctccaag agaaaggcat ctacaaagcc atgagtgagt 540

ttgacatctt catcaactac atagaagcct acatgacaat gaagatacga aactgagaca 600

tcagggtggc gactctatag actctaggac ataaattaga ggtctccaaa atcggatctg 660

gggctctggg atagctgacc cagccccttg agaaacctta ttgtacctct cttatagaat 720

atttattacc tctgatacct caacccccat ttctatttat ttactgagct tctctgtgaa 780

840

ttaagctgtt tccatagggt gacacactat ggtatttgag tgttttaaga taaattataa 900

gttacataag ggaggaaaaa aaatgttctt tggggagcca acagaagctt ccattccaag 960

cctgaccacg ctttctagct gttgagctgt ttttccctgac ctccctctaa tttatcttgt 1020

ctctggggctt ggggcttcct aactgctaca aatactctta ggaagagaaa ccagggagcc 1080

cctttgatga ttaattcacc ttccagtgtc tcggagggat tcccctaacc tcattcccca 1140

accacttcat tcttgaaagc tgtggccagc ttgttattta taacaaccta aatttggttc 1200

taggccggggc gcggtggctc acgcctgtaa tcccagcact ttgggaggct gaggcgggtg 1260

1320

ctaaaaatac aaaaattagc cgggcatggt ggcgcgcacc tgtaatccca gctacttggg 1380

aggctgaggc aagagaattg cttgaaccca ggagatggaa gttgcagtga gctgatatca 1440

tgcccctgta ctccagcctg ggtgacagag caagactctg tctcaaaaaa taaaaataaa 1500

aataaatttg gttctaatag aactcagttt taactagaat ttattcaatt cctctgggaa 1560

tgttacattg tttgtctgtc ttcatagcag attttaattt tgaataaata aatgtatctt 1620

attcacatc 1629

<210> 66

<211> 178

<212> PRT

<213> Homo sapiens

<400> 66

Met His Ser Ser Ala Leu Leu Cys Cys Leu Val Leu Leu Thr Gly Val

1 5 10 15

Arg Ala Ser Pro Gly Gln Gly Thr Gln Ser Glu Asn Ser Cys Thr His

20 25 30

Phe Pro Gly Asn Leu Pro Asn Met Leu Arg Asp Leu Arg Asp Ala Phe

35 40 45

Ser Arg Val Lys Thr Phe Phe Gln Met Lys Asp Gln Leu Asp Asn Leu

50 55 60

Leu Leu Lys Glu Ser Leu Leu Glu Asp Phe Lys Gly Tyr Leu Gly Cys

65 70 75 80

Gln Ala Leu Ser Glu Met Ile Gln Phe Tyr Leu Glu Glu Val Met Pro

85 90 95

Gln Ala Glu Asn Gln Asp Pro Asp Ile Lys Ala His Val Asn Ser Leu

100 105 110

Gly Glu Asn Leu Lys Thr Leu Arg Leu Arg Leu Arg Arg Cys His Arg

115 120 125

Phe Leu Pro Cys Glu Asn Lys Ser Lys Ala Val Glu Gln Val Lys Asn

130 135 140

Ala Phe Asn Lys Leu Gln Glu Lys Gly Ile Tyr Lys Ala Met Ser Glu

145 150 155 160

Phe Asp Ile Phe Ile Asn Tyr Ile Glu Ala Tyr Met Thr Met Lys Ile

165 170 175

Arg Asn

<210> 67

<211> 4324

<212> DNA

<213> Homo sapiens

<400> 67

cggcggcggc gagagcagag gacgagccgg gacgcggcgc cgcggcacca gggcgcgcag 60

ccgggccggc ccgaccccac cggccatacg gtggagccat cgaagccccc acccacaggc 120

tgacagaggc accgttcacc agaggggctca acaccgggat ctatgtttaa gttttaactc 180

tcgcctccaa agaccacgat aattccttcc ccaaagccca gcagcccccc agccccgcgc 240

agccccagcc tgcctcccgg cgccgatg cccgccatgc cctccagcgg ccccggggac 300

accagcagct ctgctgcgga gcgggaggag gaccgaaagg acggagagga gcaggaggag 360

420

gggaggaagc gcaagcaccc cccggtggaa agcggtgaca cgccaaagga ccctgcggtg 480

atctccaagt ccccatccat ggcccaggac tcaggcgcct cagagctatt acccaatggg 540

gacttggaga agcggagtga gccccagcca gaggagggga gccctgctgg ggggcagaag 600

ggcggggccc cagcagaggg agagggtgca gctgagaccc tgcctgaagc ctcaagagca 660

gtggaaaatg gctgctgcac ccccaaggag ggccgaggag cccctgcaga agcgggcaaa 720

gaacagaagg agaccaacat cgaatccatg aaaatggagg gctcccgggg ccggctgcgg 780

ggtggcttgg gctgggagtc cagcctccgt cagcggccca tgccgaggct caccttccag 840

gcgggggacc cctactacat cagcaagcgc aagcgggacg agtggctggc acgctggaaa 900

agggaggctg agaagaaagc caaggtcatt gcaggaatga atgctgtgga agaaaaccag 960

gggcccgggg agtctcagaa ggtggaggag gccagccctc ctgctgtgca gcagcccact 1020

gaccccgcat cccccactgt ggctaccacg cctgagcccg tggggtccga tgctggggac 1080

aagaatgcca ccaaagcagg cgatgacgag ccagagtacg aggacggccg gggctttggc 1140

attggggagc tggtgtgggg gaaactgcgg ggcttctcct ggtggccagg ccgcattgtg 1200

tcttggtgga tgacgggccg gagccgagca gctgaaggca cccgctgggt catgtggttc 1260

ggagacggca aattctcagt ggtgtgtgtt gagaagctga tgccgctgag ctcgttttgc 1320

agtgcgttcc accaggccac gtacaacaag cagcccatgt accgcaaagc catctacgag 1380

gtcctgcagg tggccagcag ccgcgcgggg aagctgttcc cggtgtgcca cgacagcgat 1440

gagagtgaca ctgccaaggc cgtggaggtg cagaacaagc ccatgattga atgggccctg 1500

gggggcttcc agccttctgg ccctaagggc ctggagccac cagaagaaga gaagaatccc 1560

tacaaagaag tgtacacgga catgtgggtg gaacctgagg cagctgccta cgcaccacct 1620

ccaccagcca aaaagccccg gaagagcaca gcggagaagc ccaaggtcaa ggagattatt 1680

gatgagcgca caagagagcg gctggtgtac gaggtgcggc agaagtgccg gaacattgag 1740

gacatctgca tctcctgtgg gagcctcaat gttaccctgg aacaccccct cttcgttgga 1800

ggaatgtgcc aaaactgcaa gaactgcttt ctggagtgtg cgtaccagta cgacgacgac 1860

ggctaccagt cctactgcac catctgctgt gggggccgtg aggtgctcat gtgcggaaac 1920

aacaactgct gcaggtgctt ttgcgtggag tgtgtggacc tcttggtggg gccgggggct 1980

gcccaggcag ccattaagga agacccctgg aactgctaca tgtgcgggca caagggtacc 2040

tacgggctgc tgcggcggcg agaggactgg ccctcccggc tccagatgtt cttcgctaat 2100

2160

aagcccatcc gggtgctgtc tctctttgat ggaatcgcta cagggctcct ggtgctgaag 2220

2280

gtgggcatgg tgcggcacca ggggaagatc atgtacgtcg gggacgtccg cagcgtcaca 2340

cagaagcata tccaggagtg gggcccattc gatctggtga ttgggggcag tccctgcaat 2400

gacctctcca tcgtcaaccc tgctcgcaag ggcctctacg agggcactgg ccggctcttc 2460

tttgagttct accgcctcct gcatgatgcg cggcccaagg agggagatga tcgccccttc 2520

ttctggctct ttgagaatgt ggtggccatg ggcgttagtg acaagaggga catctcgcga 2580

tttctcgagt ccaaccctgt gatgattgat gccaaagaag tgtcagctgc acacagggcc 2640

cgctacttct ggggtaacct tcccggtatg aacaggccgt tggcatccac tgtgaatgat 2700

aagctggagc tgcaggagtg tctggagcat ggcaggatag ccaagttcag caaagtgagg 2760

accattacta cgaggtcaaa ctccataaag cagggcaaag accagcattt tcctgtcttc 2820

atgaatgaga aagaggacat cttatggtgc actgaaatgg aaagggtatt tggtttccca 2880

gtccactata ctgacgtctc caacatgagc cgcttggcga ggcagagact gctgggccgg 2940

tcatggagcg tgccagtcat ccgccacctc ttcgctccgc tgaaggagta ttttgcgtgt 3000

3060

3120

gagaaaaagg aatttaaaac aaaaaccaca gaggcggaaa taccggaggg ctttgccttg 3180

3240

acaaaaccaa gctcccttcc cttcctcccc cttccctttt ttttcggtca gaccttttat 3300

tttctactct tttcagaggg gttttctgtt tgtttggggtt ttgtttcttg ctgtgactga 3360

3420

gaaaggtggg agagaggaaa aaaggaaatt ctatagaaat ctatatattg ggttgttttt 3480

ttttttgttt tttgtttttt ttttttgggt tttttttttt actatatatc ttttttttgt 3540

tgtctctagc ctgatcagat aggagcacaa gcaggggacg gaaagagaga gacactcagg 3600

cggcagcatt ccctcccagc cactgagctg tcgtgccagc accattcctg gtcacgcaaa 3660

acagaaccca gttagcagca gggagacgag aacaccacac aagacatttt tctacagtat 3720

3780

aaggtactgt 3840

taactactgt acaacccgac ttcataatgg tgctttcaaa cagcgagatg agtaaaaaaca 3900

3960

cttgcagatg gcgcgttctc acggtgggct cttccccttg gtttgtaacg aagtgaagga 4020

ggagaacttg ggagccaggt tctccctgcc aaaaaggggg ctagatgagg tggtcgggcc 4080

cgtggacagc tgagagtggg attcatccag actcatgcaa taaccctttg attgttttct 4140

aaaaggagac tccctcggca agatggcaga gggtacggag tcttcaggcc cagtttctca 4200

ctttagccaa ttcgagggct ccttgtggtg ggatcagaac taatccagag tgtgggaaag 4260

tgacagtcaa aaccccacct ggagcaaata aaaaaacata caaaacgtac tggtgctttc 4320

ctgt 4324

<210> 68

<211> 912

<212> PRT

<213> Homo sapiens

<400> 68

Met Pro Ala Met Pro Ser Ser Gly Pro Gly Asp Thr Ser Ser Ser Ser Ala

1 5 10 15

Ala Glu Arg Glu Glu Asp Arg Lys Asp Gly Glu Glu Gln Glu Glu Pro

20 25 30

Arg Gly Lys Glu Glu Arg Gln Glu Pro Ser Thr Thr Ala Arg Lys Val

35 40 45

Gly Arg Pro Gly Arg Lys Arg Lys His Pro Val Glu Ser Gly Asp

50 55 60

Thr Pro Lys Asp Pro Ala Val Ile Ser Lys Ser Pro Ser Met Ala Gln

65 70 75 80

Asp Ser Gly Ala Ser Glu Leu Leu Pro Asn Gly Asp Leu Glu Lys Arg

85 90 95

Ser Glu Pro Gln Pro Glu Glu Gly Ser Pro Ala Gly Gly Gln Lys Gly

100 105 110

Gly Ala Pro Ala Glu Gly Glu Gly Ala Ala Glu Thr Leu Pro Glu Ala

115 120 125

Ser Arg Ala Val Glu Asn Gly Cys Cys Thr Pro Lys Glu Gly Arg Gly

130 135 140

Ala Pro Ala Glu Ala Gly Lys Glu Gln Lys Glu Thr Asn Ile Glu Ser

145 150 155 160

Met Lys Met Glu Gly Ser Arg Gly Arg Leu Arg Gly Gly Leu Gly Trp

165 170 175

Glu Ser Ser Leu Arg Gln Arg Pro Met Pro Arg Leu Thr Phe Gln Ala

180 185 190

Gly Asp Pro Tyr Tyr Ile Ser Lys Arg Lys Arg Asp Glu Trp Leu Ala

195 200 205

Arg Trp Lys Arg Glu Ala Glu Lys Lys Ala Lys Val Ile Ala Gly Met

210 215 220

Asn Ala Val Glu Glu Asn Gln Gly Pro Gly Glu Ser Gln Lys Val Glu

225 230 235 240

Glu Ala Ser Pro Pro Ala Val Gln Gln Pro Thr Asp Pro Ala Ser Pro

245 250 255

Thr Val Ala Thr Pro Glu Pro Val Gly Ser Asp Ala Gly Asp Lys

260 265 270

Asn Ala Thr Lys Ala Gly Asp Asp Glu Pro Glu Tyr Glu Asp Gly Arg

275 280 285

Gly Phe Gly Ile Gly Glu Leu Val Trp Gly Lys Leu Arg Gly Phe Ser

290 295 300

Trp Trp Pro Gly Arg Ile Val Ser Trp Trp Met Thr Gly Arg Ser Arg

305 310 315 320

Ala Ala Glu Gly Thr Arg Trp Val Met Trp Phe Gly Asp Gly Lys Phe

325 330 335

Ser Val Val Cys Val Glu Lys Leu Met Pro Leu Ser Ser Phe Cys Ser

340 345 350

Ala Phe His Gln Ala Thr Tyr Asn Lys Gln Pro Met Tyr Arg Lys Ala

355 360 365

Ile Tyr Glu Val Leu Gln Val Ala Ser Ser Arg Ala Gly Lys Leu Phe

370 375 380

Pro Val Cys His Asp Ser Asp Glu Ser Asp Thr Ala Lys Ala Val Glu

385 390 395 400

Val Gln Asn Lys Pro Met Ile Glu Trp Ala Leu Gly Gly Phe Gln Pro

405 410 415

Ser Gly Pro Lys Gly Leu Glu Pro Pro Glu Glu Glu Lys Asn Pro Tyr

420 425 430

Lys Glu Val Tyr Thr Asp Met Trp Val Glu Pro Glu Ala Ala Ala Tyr

435 440 445

Ala Pro Pro Pro Pro Ala Lys Lys Pro Arg Lys Ser Thr Ala Glu Lys

450 455 460

Pro Lys Val Lys Glu Ile Ile Asp Glu Arg Thr Arg Glu Arg Leu Val

465 470 475 480

Tyr Glu Val Arg Gln Lys Cys Arg Asn Ile Glu Asp Ile Cys Ile Ser

485 490 495

Cys Gly Ser Leu Asn Val Thr Leu Glu His Pro Leu Phe Val Gly Gly

500 505 510

Met Cys Gln Asn Cys Lys Asn Cys Phe Leu Glu Cys Ala Tyr Gln Tyr

515 520 525

Asp Asp Asp Gly Tyr Gln Ser Tyr Cys Thr Ile Cys Cys Gly Gly Arg

530 535 540

Glu Val Leu Met Cys Gly Asn Asn Asn Cys Cys Arg Cys Phe Cys Val

545 550 555 560

Glu Cys Val Asp Leu Leu Val Gly Pro Gly Ala Ala Gln Ala Ala Ile

565 570 575

Lys Glu Asp Pro Trp Asn Cys Tyr Met Cys Gly His Lys Gly Thr Tyr

580 585 590

Gly Leu Leu Arg Arg Arg Glu Asp Trp Pro Ser Arg Leu Gln Met Phe

595 600 605

Phe Ala Asn Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro

610 615 620

Pro Val Pro Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe

625 630 635 640

Asp Gly Ile Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln

645 650 655

Val Asp Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val

660 665 670

Gly Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

675 680 685

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val

690 695 700

Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg

705 710 715 720

Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg

725 730 735

Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe

740 745 750

Trp Leu Phe Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp

755 760 765

Ile Ser Arg Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu

770 775 780

Val Ser Ala Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly

785 790 795 800

Met Asn Arg Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln

805 810 815

Glu Cys Leu Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr

820 825 830

Ile Thr Thr Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe

835 840 845

Pro Val Phe Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met

850 855 860

Glu Arg Val Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met

865 870 875 880

Ser Arg Leu Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro

885 890 895

Val Ile Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val

900 905 910

<210> 69

<211> 8789

<212> DNA

<213> Homo sapiens

<400> 69

atgctcagtg gcttctcgac aagttggcag caacaacacg gccctggtcg tcgtcgccgc 60

tgcggtaacg gagcggtttg ggtggcggag cctgcgttcg cgccttcccg ctctcctcgg 120

gaggcccttc ctgctctccc taggctccg cggccgccca gggggtggga gcgggtgagg 180

ggagccaggc gcccagcgag agaggccccc cgccgcaggg cggcccggga gctcgaggcg 240

gtccggcccg cgcgggcagc ggcgcggcgc tgaggagggg cggcctggcc gggacgcctc 300

ggggcgggggg ccgaggagct ctccggggccg ccggggaaag ctacgggccc ggtgcgtccg 360

cggaccagca gcgcgggaga gcggactccc ctcgccaccg cccgagccca ggttatcctg 420

aatacatgtc taacaatttt ccttgcaacg ttagctgttg tttttcactg tttccaaagg 480

atcaaaattg cttcagaaat tggagacata tttgatttaa aaggaaaaac ttgaacaaat 540

ggacaatatg tctattacga atacaccaac aagtaatgat gcctgtctga gcattgtgca 600

tagtttgatg tgccatagac aaggtggaga gagtgaaaca tttgcaaaaa gagcaattga 660

aagtttggta aagaagctga aggagaaaaa agatgaattg gattctttaa taacagctat 720

aactacaaat ggagctcatc ctagtaaatg tgttaccata cagagaacat tggatgggag 780

gcttcaggtg gctggtcgga aggatttcc tcatgtgatc tatgcccgtc tctggaggtg 840

gcctgatctt cacaaaaatg aactaaaaca tgttaaatat tgtcagtatg cgtttgactt 900

aaaatgtgat agtgtctgtg tgaatccata tcactacgaa cgagttgtat cacctggaat 960

tgctctctca ggattaacac tgcagagtaa tgctccatca agtatgatgg tgaaggatga 1020

atatgtgcat gactttgagg gacagccatc gttgtccact gaaggacatt caattcaaac 1080

catccagcat ccaccaagta atcgtgcatc gacagagaca tacagcaccc cagctctgtt 1140

agccccatct gagtctaatg ctaccagcac tgccaacttt cccaacattc ctgtggcttc 1200

cacaagtcag cctgccagta tactgggggg cagccatagt gaaggactgt tgcagatagc 1260

atcaggggcct cagccaggac agcagcagaa tggatttact ggtcagccag ctacttacca 1320

tcataacagc actaccacct ggactggaag taggactgca ccatacacac ctaatttgcc 1380

tcaccaccaa aacggccatc ttcagcacca cccgcctatg ccgccccatc ccggacatta 1440

ctggcctgtt cacaatgagc ttgcattcca gcctcccatt tccaatcatc ctgctcctga 1500

gtattggtgt tccattgctt actttgaaat ggatgttcag gtaggagaga catttaaggt 1560

tccttcaagc tgccctattg ttactgttga tggatacgtg gacccttctg gaggagatcg 1620

cttttgtttg ggtcaactct ccaatgtcca caggacagaa gccattgaga gagcaaggtt 1680

gcacatagggc aaaggtgtgc agttggaatg taaaggtgaa ggtgatgttt gggtcaggtg 1740

ccttagtgac cacgcggtct ttgtacagag ttactactta gacagagaag ctgggcgtgc 1800

acctggagat gctgttcata agatctaccc aagtgcatat ataaaggtct ttgatttgcg 1860

tcagtgtcat cgacagatgc agcagcaggc ggctactgca caagctgcag cagctgccca 1920

ggcagcagcc gtggcaggaa acatccctgg cccaggatca gtaggtggaa tagctccagc 1980

tatcagtctg tcagctgctg ctggaattgg tgttgatgac cttcgtcgct tatgcatact 2040

caggatgagt tttgtgaaag gctggggacc ggattaccca agacagagca tcaaagaaac 2100

accttgctgg attgaaattc acttacaccg ggccctccag ctcctagacg aagtacttca 2160

taccatgccg attgcagacc cacaaccttt agactgaggt cttttaccgt tggggccctt 2220

2280

cacttttgtt ctgctttatc ttttcataaa gggttgaaaa tgtgtttgct gccttgctcc 2340

tagcagacag aaactggatt aaaacaattt ttttttttcct cttcagaact tgtcaggcat 2400

ggctcagagc ttgaagatta ggagaaacac attcttatta attcttcacc tgttatgtat 2460

gaaggaatca ttccagtgct agaaaattta gccctttaaa acgtcttaga gccttttatc 2520

tgcagaacat cgatatgtat atcattctac agaataatcc agtattgctg attttaaagg 2580

2640

acatacttca aaaataatgt gccatgtggg tgagttaatt ttaccaagag taactttact 2700

ctgtgtttaa aaagtaagtt aataatgtat tgtaatcttt catccaaaat attttttgca 2760

agttatatta gtgaagatgg tttcaattca gattgtcttg caacttcagt tttatttttg 2820

ccaaggcaaa aaactcttaa tctgtgtgta tattgagaat cccttaaaat taccagacaa 2880

aaaaatttaa aattacgttt gttattccta gtggatgact gttgatgaag tatacttttc 2940

ccctgttaaa cagtagttgt attcttctgt atttctaggc acaaggttgg ttgctaagaa 3000

gcctataaga ggaatttctt ttccttcatt catagggaaa ggttttgtat tttttaaaac 3060

actaaaagca gcgtcactct acctaatgtc tcactgttct gcaaaggtgg caatgcttaa 3120

actaaataat gaataaactg aatattttgg aaactgctaa attctatgtt aaatactgtg 3180

3240

3300

agtttttgta tcttggggca agactgcaaa ctttttata tcttttggtt attctaagcc 3360

ctttgccatc aatgatcata tcaattggca gtgactttgt atagagaatt taagtagaaa 3420

agttgcagat gtattgactg taccacagac acaatatgta tgctttttac ctagctggta 3480

3540

ttttttttct tttgcacttt tgagtccaat ctcagtgatg aggtaccttc tactaaatga 3600

3660

3720

tactatagca gaaatagacc tgattatcta caagatgata aatagattgt ctacaggata 3780

aatagtatga aataaaatca aggattatct ttcagatgtg tttacttttg cctggagaac 3840

ttttagctat agaaacactt gtgtgatgat agtcctcctt atatcacctg gaatgaacac 3900

agcttctact gccttgctca gaaggtcttt taaatagacc atcctagaaa ccactgagtt 3960

4020

ataacttatc taccacctca tttgtactct tgattactta caaattcttt cagtaaacac 4080

ctaatttttct tctgtaaaag tttggtgatt taagttttat tggcagtttt ataaaaagac 4140

atcttctcta gaaattgcta actttaggtc cattttactg tgaatgagga ataggagtga 4200

4260

tgacataatt cattgcttct ttttttttgag atatggagtc ttgctgtgtt gcccaggcag 4320

gagtgcagtg gtatgatctc agctcactgc aacctctgcc tcccgggttc aactgattct 4380

cctgcctcag cctccctggt agctaggatt acaggtgccc gccaccatgc ctggctaact 4440

tttgtagttt tagtagagac ggggttttgc ctgttggcca ggctggtctt gaactcctga 4500

cctcaagtga tccatccacc ttggcctccc aaagtgctgg gattacgggc gtgagccact 4560

gtccctggcc tcattgttcc cttttctact ttaaggaaag ttttcatgtt taatcatctg 4620

gggaaagtat gtgaaaaata tttgttaaga agtatctctt tggagccaag ccacctgtct 4680

tggtttcttt ctactaagag ccataaagta tagaaatact tctagttgtt aagtgcttat 4740

atttgtacct agatttagtc acacgctttt gagaaaacat ctagtatgtt atgatcagct 4800

attcctgaga gcttggttgt taatctatat ttctatttct tagtggtagt catctttgat 4860

gaataagact aaagattctc acaggtttaa aattttatgt ctactttaag ggtaaaatta 4920

tgaggttatg gttctgggtg ggttttctct agctaattca tatctcaaag agtctcaaaa 4980

tgttgaattt cagtgcaagc tgaatgagag atgagccatg tacacccacc gtaagacctc 5040

attccatgtt tgtccagtgc ctttcagtgc attatcaaag ggaatccttc atggtgttgc 5100

ctttattttc cggggagtag atcgtgggat atagtctatc tcatttttaa tagtttaccg 5160

cccctggtat acaaagataa tgacaataaa tcactgccat ataaccttgc tttttccaga 5220

aacatggctg ttttgtattg ctgtaaccac taaataggtt gcctatacca ttcctcctgt 5280

5340

tgagtaaact gaactcatat tagctgtgct gcatttcaga cttaaaatcc atttttgtgg 5400

ggcagggtgt ggtgtgtaaa ggggggtgtt tgtaatacaa gttgaaggca aaataaaatg 5460

tcctgtctcc cagatgatat acatcttatt atttttaaag tttattgcta attgtaggaa 5520

ggtgagttgc aggtatcttt gactatggtc atctggggaa ggaaaatttt acattttact 5580

attaatgctc cttaagtgtc tatggaggtt aaagaataaa atggtaaatg tttctgtgcc 5640

tggtttgatg gtaactggtt aatagttact caccatttta tgcagagtca cattagttca 5700

caccctttct gagagccttt tgggagaagc agttttattc tctgagtgga acagagttct 5760

ttttgttgat aatttctagt ttgctccctt cgttattgcc aactttactg gcattttatt 5820

taatgatagc agattgggaa aatggcaaat ttaggttacg gaggtaaatg agtatatgaa 5880

agcaattacc tctaaagcca gttaacaatt attttgtagg tggggtacac tcagcttaaa 5940

gtaatgcatt ttttttttccc gtaaaggcag aatccatctt gttgcagata gctatctaaa 6000

taatctcata tcctcttttg caaagactac agagaatagg ctatgacaat cttgttcaag 6060

6120

aaaagtccat ggccttattc atccacaaag tggcatccta ggcccagcct tatccctagc 6180

agttgtccca gtgctgctag gttgcttatc ttgtttatct ggaatcactg tggagtgaaa 6240

ttttccacat catccagaat tgccttattt aagaagtaaa acgttttaat ttttagcctt 6300

6360

ctgtgcttgg ctatctttgt ggacttcagg ggcttctaaa acagacagga ctgtgttgcc 6420

tttactaaat ggtctgagac agctatggtt ttgaattttt agtttttttt ttttaaccca 6480

cttcccctcc tggtctcttc cctctctgat aattaccatt catatgtgag tgttagtgtg 6540

cctcctttta gcattttctt cttctctttc tgattcttca ttttctgactg cctaggcaag 6600

aaccaaactt actagaacgt tctttaaaac acaagtacaa actctgggac 6660

aggacccaag acactttcct gtgaagtgct gaaaaagacc tcattgtatt ggcatttgat 6720

atcagtttga tgtagcttag agtgcttcct gattcttgct gagtttcagg tagttgagat 6780

agagagaagt gagtcatatt catatttttcc cccttagaat aatattttga aaggtttcat 6840

tgcttccact tgaatgctgc tcttacaaaa actggggtta caagggttac taaattagca 6900

tcagtagcca gaggcaatac cgttgtctgg aggacaccag caaacaacac acaacaaagc 6960

aaaacaaacc ttgggaaact aaggccattt gttttgtttt ggtgtcccct ttgaagccct 7020

gccttctggc cttactcctg tacagatatt tttgacctat aggtgccttt atgagaattg 7080

agggtctgac atcctgcccc aaggagtagc taaagtaatt gctagtgttt tcagggattt 7140

taacatcaga ctggaatgaa tgaatgaaac tttttgtcct ttttttttct gttttttttt 7200

ttctaatgta gtaaggacta aggaaaacct ttggtgaaga caatcatttc tctctgttga 7260

tgtggatact tttcacaccg tttatttaaa tgctttctca ataggtccag agccagtgtt 7320

cttgttcaac ctgaaagtaa tggctctggg ttgggccaga cagttgcact ctctagtttg 7380

ccctctgcca caaatttgat gtgtgacctt tgggcaagtc atttatcttc tctgggcctt 7440

agttgcctca tctgtaaaat gagggagttg gagtagatta attattccag ctctgaaatt 7500

ctaagtgacc ttggctacct tgcagcagtt ttggatttct tccttatctt tgttctgctg 7560

tttgagggggg ctttttactt atttccatgt tattcaaagg agactaggct tgatatttta 7620

ttactgttct tttatggaca aaaggttaca tagtatgccc ttaagactta attttaacca 7680

aaggcctagc accaccttag gggctgcaat aaacacttaa cgcgcgtgcg cacgcgcgcg 7740

7800

tgacattcta gcttttgaat tgcgtgcaca cacacacgca cgcacacact ctggtcagag 7860

tttattaagg ctttcgagtc atgacattat agcttttgag ttggtgtgtg tgacaccacc 7920

ctcctaagtg gtgtgtgctt gtaatttttt ttttcagtga aaatggattg aaaacctgtt 7980

gttaatgctt agtgatatta tgctcaaaac aaggaaattc ccttgaaccg tgtcaattaa 8040

actggtttat atgactcaag aaaacaatac cagtagatga ttattaactt tattcttggc 8100

tctttttagg tccattttga ttaagtgact tttggctgga tcattcagag ctctcttcta 8160

gcctaccctt ggatgagtac aattaatgaa attcatattt tcaaggacct gggagccttc 8220

cttggggctg ggttgagggt ggggggttgg ggagtcctgg tagaggccag ctttgtggta 8280

gctggagagg aagggatgaa accagctgct gttgcaaagg ctgcttgtca ttgatagaag 8340

gactcacggg cttggattga ttaagactaa acatggagtt ggcaaacttt cttcaagtat 8400

tgagttctgt tcaatgcatt ggacatgtga tttaagggaa aagtgtgaat gcttatagat 8460

gatgaaaacc tggtgggctg cagagcccag tttagaagaa gtgagttggg ggttggggac 8520

agatttggtg gtggtatttc ccaactgttt cctcccctaa attcagagga atgcagctat 8580

gccagaagcc agagaagagc cactcgtagc ttctgctttg gggacaactg gtcagttgaa 8640

agtccgga gttcctttgt ggctttctgt atacttttgc ctggttaaag tctgtggcta 8700

aaaaatagtc gaacctttct tgagaactct gtaacaaagt atgtttttga ttaaaagaga 8760

aagccaacta 8789

<210> 70

<211> 552

<212> PRT

<213> Homo sapiens

<400> 70

Met Asp Asn Met Ser Ile Thr Asn Thr Pro Thr Ser Asn Asp Ala Cys

1 5 10 15

Leu Ser Ile Val His Ser Leu Met Cys His Arg Gln Gly Gly Glu Ser

20 25 30

Glu Thr Phe Ala Lys Arg Ala Ile Glu Ser Leu Val Lys Lys Leu Lys

35 40 45

Glu Lys Lys Asp Glu Leu Asp Ser Leu Ile Thr Ala Ile Thr Asn

50 55 60

Gly Ala His Pro Ser Lys Cys Val Thr Ile Gln Arg Thr Leu Asp Gly

65 70 75 80

Arg Leu Gln Val Ala Gly Arg Lys Gly Phe Pro His Val Ile Tyr Ala

85 90 95

Arg Leu Trp Arg Trp Pro Asp Leu His Lys Asn Glu Leu Lys His Val

100 105 110

Lys Tyr Cys Gln Tyr Ala Phe Asp Leu Lys Cys Asp Ser Val Cys Val

115 120 125

Asn Pro Tyr His Tyr Glu Arg Val Val Ser Pro Gly Ile Asp Leu Ser

130 135 140

Gly Leu Thr Leu Gln Ser Asn Ala Pro Ser Ser Met Met Val Lys Asp

145 150 155 160

Glu Tyr Val His Asp Phe Glu Gly Gln Pro Ser Leu Ser Thr Glu Gly

165 170 175

His Ser Ile Gln Thr Ile Gln His Pro Pro Ser Asn Arg Ala Ser Thr

180 185 190

Glu Thr Tyr Ser Thr Pro Ala Leu Leu Ala Pro Ser Glu Ser Asn Ala

195 200 205

Thr Ser Thr Ala Asn Phe Pro Asn Ile Pro Val Ala Ser Thr Ser Gln

210 215 220

Pro Ala Ser Ile Leu Gly Gly Ser His Ser Glu Gly Leu Leu Gln Ile

225 230 235 240

Ala Ser Gly Pro Gln Pro Gly Gln Gln Gln Asn Gly Phe Thr Gly Gln

245 250 255

Pro Ala Thr Tyr His His Asn Ser Thr Thr Thr Thr Trp Thr Gly Ser Arg

260 265 270

Thr Ala Pro Tyr Thr Pro Asn Leu Pro His His Gln Asn Gly His Leu

275 280 285

Gln His His Pro Pro Met Pro Pro His Pro Gly His Tyr Trp Pro Val

290 295 300

His Asn Glu Leu Ala Phe Gln Pro Pro Ile Ser Asn His Pro Ala Pro

305 310 315 320

Glu Tyr Trp Cys Ser Ile Ala Tyr Phe Glu Met Asp Val Gln Val Gly

325 330 335

Glu Thr Phe Lys Val Pro Ser Ser Cys Pro Ile Val Thr Val Asp Gly

340 345 350

Tyr Val Asp Pro Ser Gly Gly Asp Arg Phe Cys Leu Gly Gln Leu Ser

355 360 365

Asn Val His Arg Thr Glu Ala Ile Glu Arg Ala Arg Leu His Ile Gly

370 375 380

Lys Gly Val Gln Leu Glu Cys Lys Gly Glu Gly Asp Val Trp Val Arg

385 390 395 400

Cys Leu Ser Asp His Ala Val Phe Val Gln Ser Tyr Tyr Leu Asp Arg

405 410 415

Glu Ala Gly Arg Ala Pro Gly Asp Ala Val His Lys Ile Tyr Pro Ser

420 425 430

Ala Tyr Ile Lys Val Phe Asp Leu Arg Gln Cys His Arg Gln Met Gln

435 440 445

Gln Gln Ala Ala Thr Ala Gln Ala Ala Ala Ala Ala Gln Ala Ala Ala

450 455 460

Val Ala Gly Asn Ile Pro Gly Pro Gly Ser Val Gly Gly Ile Ala Pro

465 470 475 480

Ala Ile Ser Leu Ser Ala Ala Ala Gly Ile Gly Val Asp Asp Leu Arg

485 490 495

Arg Leu Cys Ile Leu Arg Met Ser Phe Val Lys Gly Trp Gly Pro Asp

500 505 510

Tyr Pro Arg Gln Ser Ile Lys Glu Thr Pro Cys Trp Ile Glu Ile His

515 520 525

Leu His Arg Ala Leu Gln Leu Leu Asp Glu Val Leu His Thr Met Pro

530 535 540

Ile Ala Asp Pro Gln Pro Leu Asp

545 550

<210> 71

<211> 5477

<212> DNA

<213> Homo sapiens

<400> 71

aaaaagagaa actgttggga gaggaatcgt atctccatat ttcttctttc agccccaatc 60

caagggttgt agctggaact ttccatcagt tcttcctttc ttttcctct ctaagccttt 120

gccttgctct gtcacagtga agtcagccag agcagggctg ttaaactctg tgaaatttgt 180

cataagggtg tcaggtattt cttactggct tccaaagaaa catagataaa gaaatctttc 240

ctgtggcttc ccttggcagg ctgcattcag aaggtctctc agttgaagaa agagcttgga 300

ggacaacagc acaacaggag agtaaaagat gccccagggc tgaggcctcc gctcaggcag 360

ccgcatctgg ggtcaatcat actcaccttg ccggggccat gctccagcaa aatcaagctg 420

ttttcttttg aaagttcaaa ctcatcaaga ttatgctgct cactcttatc attctgttgc 480

cagtagtttc aaaatttagt tttgttagtc tctcagcacc gcagcactgg agctgtcctg 540

aaggtactct cgcaggaaat gggaattcta cttgtgtggg tcctgcaccc ttcttaattt 600

tctcccatgg aaatagtatc tttaggattg aacacagaagg aaccaattat gagcaattgg 660

tggtggatgc tggtgtctca gtgatcatgg attttcatta taatgagaaa agaatctatt 720

gggtggattt agaaagacaa cttttgcaaa gagtttttct gaatggggtca aggcaagaga 780

gagtatgtaa tatagagaaa aatgtttctg gaatggcaat aaattggata aatgaagaag 840

ttatttggtc aaatcaacag gaaggaatca ttacagtaac agatatgaaa ggaaataatt 900

cccacattct tttaagtgct ttaaaatatc ctgcaaatgt agcagttgat ccagtagaaa 960

ggtttatatt ttggtcttca gaggtggctg gaagccttta tagagcagat ctcgatggtg 1020

tgggagtgaa ggctctgttg gagacatcag agaaaataac agctgtgtca ttggatgtgc 1080

ttgataagcg gctgttttgg attcagtaca acagagaagg aagcaattct cttatttgct 1140

cctgtgatta tgatggaggt tctgtccaca ttagtaaaca tccaacacag cataatttgt 1200

ttgcaatgtc ccttttttggt gaccgtatct tctattcaac atggaaaatg aagacaattt 1260

ggatagccaa caaacacact ggaaaggaca tggttagaat taacctccat tcatcatttg 1320

taccacttgg tgaactgaaa gtagtgcatc cacttgcaca acccaaggca gaagatgaca 1380

cttgggagcc tgagcagaaa ctttgcaaat tgaggaaagg aaactgcagc agcactgtgt 1440

gtgggcaaga cctccagtca cacttgtgca tgtgtgcaga gggatacgcc ctaagtcgag 1500

accggaagta ctgtgaagat gttaatgaat gtgctttttg gaatcatggc tgtactcttg 1560

ggtgtaaaaa cacccctgga tcctattact gcacgtgccc tgtaggattt gttctgcttc 1620

ctgatgggaa acgatgtcat caacttgttt cctgtccacg caatgtgtct gaatgcagcc 1680

atgactgtgt tctgacatca gaaggtccct tatgtttctg tcctgaaggc tcagtgcttg 1740

agagagatgg gaaaacatgt agcggttgtt cctcacccga taatggtgga tgtagccagc 1800

tctgcgttcc tcttagccca gtatcctggg aatgtgattg ctttcctggg tatgacctac 1860

aactggatga aaaaagctgt gcagcttcag gaccacaacc atttttgctg tttgccaatt 1920

ctcaagatat tcgacacatg cattttgatg gaacagacta tggaactctg ctcagccagc 1980

agatgggaat ggtttatgcc ctagatcatg accctgtgga aaataagata tactttgccc 2040

atacagccct gaagtggata gagagagcta atatggatgg ttcccagcga gaaaggctta 2100

ttgaggaagg agtagatgtg ccagaaggtc ttgctgtgga ctggattggc cgtagattct 2160

attggacaga cagagggaaa tctctgattg gaaggagtga tttaaatgggg aaacgttcca 2220

aaataatcac taaggagaac atctctcaac cacgaggaat tgctgttcat ccaatggcca 2280

agagattatt ctggactgat acagggatta atccacgaat tgaaagttct tccctccaag 2340

gccttggccg tctggttata gccagctctg atctaatctg gcccagtgga ataacgattg 2400

acttcttaac tgacaagttg tactggtgcg atgccaagca gtctgtgatt gaaatggcca 2460

atctggatgg ttcaaaacgc cgaagactta cccagaatga tgtaggtcac ccatttgctg 2520

tagcagtgtt tgaggattat gtgtggttct cagattgggc tatgccatca gtaatgagag 2580

taaacaagag gactggcaaa gatagagtac gtctccaagg cagcatgctg aagccctcat 2640

cactggttgt ggttcatcca ttggcaaaac caggagcaga tccctgctta tatcaaaacg 2700

2760

gttttatgaa agcctcagat gggaaaacgt gtctggctct ggatggtcat cagctgttgg 2820

caggtggtga agttgatcta aagaaccaag taacaccatt ggacatcttg tccaagacta 2880

gagtgtcaga agataacatt acagaatctc aacacatgct agtggctgaa atcatggtgt 2940

cagatcaaga tgactgtgct cctgtgggat gcagcatgta tgctcggtgt atttcagagg 3000

3060

atatagatga atgtgagatg ggtgtcccag tgtgcccccc tgcctcctcc aagtgcatca 3120

acaccgaagg tggttatgtc tgccggtgct cagaaggcta ccaaggagat gggattcact 3180

gtcttgactc tactccaccc cctcacctca gggaagatga ccaccactat tccgtaagaa 3240

atagtgactc tgaatgtccc ctgtcccacg atgggtactg cctccatgat ggtgtgtgca 3300

tgtatattga agcattggac aagtatgcat gcaactgtgt tgttggctac atcggggagc 3360

gatgtcagta ccgagacctg aagtggtggg aactgcgcca cgctggccac gggcagcagc 3420

agaaggtcat cgtggtggct gtctgcgtgg tggtgcttgt catgctgctc ctcctgagcc 3480

tgtggggggc ccactactac aggactcaga agctgctatc gaaaaaccca aagaatcctt 3540

atgaggagtc gagcagagat gtgaggagtc gcaggcctgc tgacactgag gatgggatgt 3600

cctcttgccc tcaaccttgg tttgtggtta taaaagaaca ccaagacctc aagaatgggg 3660

gtcaaccagt ggctggtgag gatggccagg cagcagatgg gtcaatgcaa ccaacttcat 3720

ggaggcagga gccccagtta tgtggaatgg gcacagagca aggctgctgg attccagtat 3780

ccagtgataa gggctcctgt ccccaggtaa tggagcgaag ctttcatatg ccctcctatg 3840

ggacacagac ccttgaaggg ggtgtcgaga agccccattc tctcctatca gctaacccat 3900

tatggcaaca aagggccctg gacccaccac accaaatgga gctgactcag tgaaaactgg 3960

aattaaaagg aaagtcaaga agaatgaact atgtcgatgc acagtatctt ttctttcaaa 4020

agtagagcaa aactataggt tttggttcca caatctctac gactaatcac ctactcaatg 4080

cctggagaca gatacgtagt tgtgcttttg tttgctcttt taagcagtct cactgcagtc 4140

ttatttccaa gtaagagtac tgggagaatc actaggtaac ttattagaaa cccaaattgg 4200

4260

ttgttgttcc tgcagcccca gaagaaatta ggggttaaag cagacagtca cactggtttg 4320

gtcagttaca aagtaatttc tttgatctgg acagaacatt tatatcagtt tcatgaaatg 4380

attggaatat tacaataccg ttaagataca gtgtaggcat ttaactcctc attggcgtgg 4440

tccatgctga tgattttgca aaatgagttg tgatgaatca atgaaaaatg taatttagaa 4500

actgatttct tcagaattag atggcttatt ttttaaaata tttgaatgaa aacattttat 4560

ttttaaaata ttacacagga ggcttcggag ttttcttagtc attactgtcc ttttccccta 4620

cagaattttc cctcttggtg tgattgcaca gaatttgtat gtattttcag ttacaagatt 4680

gtaagtaaat tgcctgattt gttttcatta tagacaacga tgaatttctt ctaattattt 4740

aaataaaatc accaaaaaca taaacatttt attgtatgcc tgattaagta gttaattata 4800

4860

tttcgtttt ttttttttttc cggagagagg ataggatctc actctgttat ccaggctgga 4920

gtgtgcaatg gcacaatcat agctcagtgc agcctcaaac tcctgggctc aagcaatcct 4980

cctgcctcag cctcccgagt aactaggacc acaggcacag gccaccatgc ctggctaagg 5040

tttttatttt tattttttgt agacatgggg atcacacaat gttgcccagg ctggtcttga 5100

actcctggcc tcaagcaagg tcgtgctggt aattttgcaa aatgaattgt gattgacttt 5160

cagcctccca acgtattaga ttataggcat tagccatggt gcccagcctt gtaactttta 5220

aaaaaatttt ttaatctaca actctgtaga ttaaaatttc acatggtgtt ctaattaaat 5280

atttttcttg cagccaagat attgttacta cagataacac aacctgatat ggtaacttta 5340

aattttgggg gctttgaatc attcagttta tgcattaact agtccctttg tttatctttc 5400

atttctcaac cccttgtact ttggtgatac cagacatcag aataaaaaga aattgaagta 5460

aaaaaaaaaaaaaaaaa 5477

<210> 72

<211> 1166

<212> PRT

<213> Homo sapiens

<400> 72

Met Leu Leu Thr Leu Ile Ile Leu Leu Pro Val Val Ser Lys Phe Ser

1 5 10 15

Phe Val Ser Leu Ser Ala Pro Gln His Trp Ser Cys Pro Glu Gly Thr

20 25 30

Leu Ala Gly Asn Gly Asn Ser Thr Cys Val Gly Pro Ala Pro Phe Leu

35 40 45

Ile Phe Ser His Gly Asn Ser Ile Phe Arg Ile Asp Thr Glu Gly Thr

50 55 60

Asn Tyr Glu Gln Leu Val Val Asp Ala Gly Val Ser Val Ile Met Asp

65 70 75 80

Phe His Tyr Asn Glu Lys Arg Ile Tyr Trp Val Asp Leu Glu Arg Gln

85 90 95

Leu Leu Gln Arg Val Phe Leu Asn Gly Ser Arg Gln Glu Arg Val Cys

100 105 110

Asn Ile Glu Lys Asn Val Ser Gly Met Ala Ile Asn Trp Ile Asn Glu

115 120 125

Glu Val Ile Trp Ser Asn Gln Gln Glu Gly Ile Ile Thr Val Thr Asp

130 135 140

Met Lys Gly Asn Asn Ser His Ile Leu Leu Ser Ala Leu Lys Tyr Pro

145 150 155 160

Ala Asn Val Ala Val Asp Pro Val Glu Arg Phe Ile Phe Trp Ser Ser

165 170 175

Glu Val Ala Gly Ser Leu Tyr Arg Ala Asp Leu Asp Gly Val Gly Val

180 185 190

Lys Ala Leu Leu Glu Thr Ser Glu Lys Ile Thr Ala Val Ser Leu Asp

195 200 205

Val Leu Asp Lys Arg Leu Phe Trp Ile Gln Tyr Asn Arg Glu Gly Ser

210 215 220

Asn Ser Leu Ile Cys Ser Cys Asp Tyr Asp Gly Gly Ser Val His Ile

225 230 235 240

Ser Lys His Pro Thr Gln His Asn Leu Phe Ala Met Ser Leu Phe Gly

245 250 255

Asp Arg Ile Phe Tyr Ser Thr Trp Lys Met Lys Thr Ile Trp Ile Ala

260 265 270

Asn Lys His Thr Gly Lys Asp Met Val Arg Ile Asn Leu His Ser Ser

275 280 285

Phe Val Pro Leu Gly Glu Leu Lys Val Val His Pro Leu Ala Gln Pro

290 295 300

Lys Ala Glu Asp Asp Thr Trp Glu Pro Glu Gln Lys Leu Cys Lys Leu

305 310 315 320

Arg Lys Gly Asn Cys Ser Ser Thr Val Cys Gly Gln Asp Leu Gln Ser

325 330 335

His Leu Cys Met Cys Ala Glu Gly Tyr Ala Leu Ser Arg Asp Arg Lys

340 345 350

Tyr Cys Glu Asp Val Asn Glu Cys Ala Phe Trp Asn His Gly Cys Thr

355 360 365

Leu Gly Cys Lys Asn Thr Pro Gly Ser Tyr Tyr Cys Thr Cys Pro Val

370 375 380

Gly Phe Val Leu Leu Pro Asp Gly Lys Arg Cys His Gln Leu Val Ser

385 390 395 400

Cys Pro Arg Asn Val Ser Glu Cys Ser His Asp Cys Val Leu Thr Ser

405 410 415

Glu Gly Pro Leu Cys Phe Cys Pro Glu Gly Ser Val Leu Glu Arg Asp

420 425 430

Gly Lys Thr Cys Ser Gly Cys Ser Ser Pro Asp Asn Gly Gly Cys Ser

435 440 445

Gln Leu Cys Val Pro Leu Ser Pro Val Ser Trp Glu Cys Asp Cys Phe

450 455 460

Pro Gly Tyr Asp Leu Gln Leu Asp Glu Lys Ser Cys Ala Ala Ser Gly

465 470 475 480

Pro Gln Pro Phe Leu Leu Phe Ala Asn Ser Gln Asp Ile Arg His Met

485 490 495

His Phe Asp Gly Thr Asp Tyr Gly Thr Leu Leu Ser Gln Gln Met Gly

500 505 510

Met Val Tyr Ala Leu Asp His Asp Pro Val Glu Asn Lys Ile Tyr Phe

515 520 525

Ala His Thr Ala Leu Lys Trp Ile Glu Arg Ala Asn Met Asp Gly Ser

530 535 540

Gln Arg Glu Arg Leu Ile Glu Glu Gly Val Asp Val Pro Glu Gly Leu

545 550 555 560

Ala Val Asp Trp Ile Gly Arg Arg Phe Tyr Trp Thr Asp Arg Gly Lys

565 570 575

Ser Leu Ile Gly Arg Ser Asp Leu Asn Gly Lys Arg Ser Lys Ile Ile

580 585 590

Thr Lys Glu Asn Ile Ser Gln Pro Arg Gly Ile Ala Val His Pro Met

595 600 605

Ala Lys Arg Leu Phe Trp Thr Asp Thr Gly Ile Asn Pro Arg Ile Glu

610 615 620

Ser Ser Ser Leu Gln Gly Leu Gly Arg Leu Val Ile Ala Ser Ser Asp

625 630 635 640

Leu Ile Trp Pro Ser Gly Ile Thr Ile Asp Phe Leu Thr Asp Lys Leu

645 650 655

Tyr Trp Cys Asp Ala Lys Gln Ser Val Ile Glu Met Ala Asn Leu Asp

660 665 670

Gly Ser Lys Arg Arg Arg Leu Thr Gln Asn Asp Val Gly His Pro Phe

675 680 685

Ala Val Ala Val Phe Glu Asp Tyr Val Trp Phe Ser Asp Trp Ala Met

690 695 700

Pro Ser Val Met Arg Val Asn Lys Arg Thr Gly Lys Asp Arg Val Arg

705 710 715 720

Leu Gln Gly Ser Met Leu Lys Pro Ser Ser Leu Val Val Val His Pro

725 730 735

Leu Ala Lys Pro Gly Ala Asp Pro Cys Leu Tyr Gln Asn Gly Gly Cys

740 745 750

Glu His Ile Cys Lys Lys Arg Leu Gly Thr Ala Trp Cys Ser Cys Arg

755 760 765

Glu Gly Phe Met Lys Ala Ser Asp Gly Lys Thr Cys Leu Ala Leu Asp

770 775 780

Gly His Gln Leu Leu Ala Gly Gly Glu Val Asp Leu Lys Asn Gln Val

785 790 795 800

Thr Pro Leu Asp Ile Leu Ser Lys Thr Arg Val Ser Glu Asp Asn Ile

805 810 815

Thr Glu Ser Gln His Met Leu Val Ala Glu Ile Met Val Ser Asp Gln

820 825 830

Asp Asp Cys Ala Pro Val Gly Cys Ser Met Tyr Ala Arg Cys Ile Ser

835 840 845

Glu Gly Glu Asp Ala Thr Cys Gln Cys Leu Lys Gly Phe Ala Gly Asp

850 855 860

Gly Lys Leu Cys Ser Asp Ile Asp Glu Cys Glu Met Gly Val Pro Val

865 870 875 880

Cys Pro Pro Ala Ser Ser Lys Cys Ile Asn Thr Glu Gly Gly Tyr Val

885 890 895

Cys Arg Cys Ser Glu Gly Tyr Gln Gly Asp Gly Ile His Cys Leu Asp

900 905 910

Ser Thr Pro Pro Pro His Leu Arg Glu Asp Asp His His Tyr Ser Val

915 920 925

Arg Asn Ser Asp Ser Glu Cys Pro Leu Ser His Asp Gly Tyr Cys Leu

930 935 940

His Asp Gly Val Cys Met Tyr Ile Glu Ala Leu Asp Lys Tyr Ala Cys

945 950 955 960

Asn Cys Val Val Gly Tyr Ile Gly Glu Arg Cys Gln Tyr Arg Asp Leu

965 970 975

Lys Trp Trp Glu Leu Arg His Ala Gly His Gly Gln Gln Gln Lys Val

980 985 990

Ile Val Val Ala Val Cys Val Val Val Leu Val Met Leu Leu Leu Leu

995 1000 1005

Ser Leu Trp Gly Ala His Tyr Tyr Arg Thr Gln Lys Leu Leu Ser

1010 1015 1020

Lys Asn Pro Lys Asn Pro Tyr Glu Glu Ser Ser Arg Asp Val Arg

1025 1030 1035

Ser Arg Arg Pro Ala Asp Thr Glu Asp Gly Met Ser Ser Cys Pro

1040 1045 1050

Gln Pro Trp Phe Val Val Ile Lys Glu His Gln Asp Leu Lys Asn

1055 1060 1065

Gly Gly Gln Pro Val Ala Gly Glu Asp Gly Glyn Ala Ala Asp Gly

1070 1075 1080

Ser Met Gln Pro Thr Ser Trp Arg Gln Glu Pro Gln Leu Cys Gly

1085 1090 1095

Met Gly Thr Glu Gln Gly Cys Trp Ile Pro Val Ser Ser Asp Lys

1100 1105 1110

Gly Ser Cys Pro Gln Val Met Glu Arg Ser Phe His Met Pro Ser

1115 1120 1125

Tyr Gly Thr Gln Thr Leu Glu Gly Gly Val Glu Lys Pro His Ser

1130 1135 1140

Leu Leu Ser Ala Asn Pro Leu Trp Gln Gln Arg Ala Leu Asp Pro

1145 1150 1155

Pro His Gln Met Glu Leu Thr Gln

1160 1165

<210> 73

<211> 4815

<212> DNA

<213> Homo sapiens

<400> 73

agtggcgtcg gaactgcaaa gcacctgtga gcttgcggaa gtcagttcag actccagccc 60

gctccagccc ggcccgaccc gaccgcaccc ggcgcctgcc ctcgctcggc gtccccggcc 120

agccatgggc ccttggagcc gcagcctctc ggcgctgctg ctgctgctgc aggtctcctc 180

ttggctctgc caggagccgg agccctgcca ccctggcttt gacgccgaga gctacacgtt 240

cacggtgccc cggcgccacc tggagagagg ccgcgtcctg ggcagagtga attttgaaga 300

ttgcaccggt cgacaaagga cagcctattt ttccctcgac acccgattca aagtgggcac 360

agatggtgtg attacagtca aaaggcctct acggtttcat aacccacaga tccatttctt 420

ggtctacgcc tgggactcca cctacagaaa gttttccacc aaagtcacgc tgaatacagt 480

ggggcaccac caccgccccc cgccccatca ggcctccgtt tctggaatcc aagcagaatt 540

gctcacattt cccaactcct ctcctggcct cagaagacag aagagagact gggttattcc 600

tcccatcagc tgccgagaaa atgaaaaagg cccatttcct aaaaacctgg ttcagatcaa 660

atccaacaaa gacaaagaag gcaaggtttt ctacagcatc actggccaag gagctgacac 720

accccctgtt ggtgtcttta ttattgaaag agaaacagga tggctgaagg tgacagagcc 780

tctggataga gaacgcattg ccacatacac tctcttctct cacgctgtgt catccaacgg 840

gaatgcagtt gaggatccaa tggagatttt gatcacggta accgatcaga atgacaacaa 900

gcccgaattc acccaggagg tctttaaggg gtctgtcatg gaaggtgctc ttcggaac 960

ctctgtgatg gaggtcacag ccacagacgc ggacgatgat gtgaacacct acaatgccgc 1020

catcgcttac accatcctca gccaagatcc tgagctccct gacaaaaata tgttcaccat 1080

taacaggaac acaggagtca tcagtgtggt caccactggg ctggaccgag agagtttccc 1140

tacgtatacc ctggtggttc aagctgctga ccttcaaggt gaggggttaa gcacaacagc 1200

aacagctgtg atcacagtca ctgacaccaa cgataatcct ccgatcttca atcccaccac 1260

gtacaagggt caggtgcctg agaacgaggc taacgtcgta atcaccacac tgaaagtgac 1320

tgatgctgat gcccccaata ccccagcgtg ggaggctgta tacaccatat tgaatgatga 1380

1440

aaagggcttg gattttgagg ccaagcagca gtacattcta cacgtagcag tgacgaatgt 1500

ggtacctttt gaggtctctc tcaccacctc cacagccacc gtcaccgtgg atgtgctgga 1560

tgtgaatgaa gcccccatct ttgtgcctcc tgaaaagaga gtggaagtgt ccgaggactt 1620

tggcgtgggc caggaaatca catcctacac tgcccaggag ccagacacat ttatggaaca 1680

gaaaataaca tatcggattt ggagagacac tgccaactgg ctggagatta atccggacac 1740

1800

cacgtacaca gccctaatca tagctacaga caatggttct ccagttgcta ctggaacagg 1860

gacacttctg ctgatcctgt ctgatgtgaa tgacaacgcc cccataccag aacctcgaac 1920

tatattcttc tgtgagagga atccaaagcc tcaggtcata aacatcattg atgcagacct 1980

tcctcccaat acatctccct tcacagcaga actaacacac ggggcgagtg ccaactggac 2040

cattcagtac aacgacccaa cccaagaatc tatcattttg aagccaaaga tggccttaga 2100

ggtgggtgac tacaaaatca atctcaagct catggataac cagaataaag accaagtgac 2160

caccttagag gtcagcgtgt gtgactgtga aggggccgct ggcgtctgta ggaaggcaca 2220

gcctgtcgaa gcaggattgc aaattcctgc cattctgggg attcttggag gaattcttgc 2280

tttgctaatt ctgattctgc tgctcttgct gtttcttcgg aggagagcgg tggtcaaaga 2340

gcccttactg cccccagagg atgacacccg ggacaacgtt tattactatg atgaagaagg 2400

aggcggagaa gaggaccagg actttgactt gagccagctg cacaggggcc tggacgctcg 2460

gcctgaagtg actcgtaacg acgttgcacc aaccctcatg agtgtccccc ggtatcttcc 2520

ccgccctgcc aatcccgatg aaattggaaa tttttattgat gaaaatctga aagcggctga 2580

tactgacccc acagccccgc cttatgattc tctgctcgtg tttgactatg aaggaagcgg 2640

ttccgaagct gctagtctga gctccctgaa ctcctcagag tcagacaaag accaggacta 2700

tgactacttg aacgaatggg gcaatcgctt caagaagctg gctgacatgt acggaggcgg 2760

cgaggacgac taggggactc gagagaggcg ggccccagac ccatgtgctg ggaaatgcag 2820

2880

aaagagactg gttagtgatg cagttagtat agctttatac tctctccact ttatagctct 2940

aataagtttg tgttagaaaa gtttcgactt atttcttaaa gctttttttt ttttcccatc 3000

actctttaca tggtggtgat gtccaaaaga tacccaaatt ttaatattcc agaagaacaa 3060

3120

ttttaaaaag aaggggagaa gtcagctact ctagttctgt tgttttgtgt atataatttt 3180

ttaaaaaaaa tttgtgtgct tctgctcatt actacactgg tgtgtccctc tgcctttttt 3240

ttttttttaa gacagggtct cattctatcg gccaggctgg agtgcagtgg tgcaatcaca 3300

gctcactgca gccttgtcct cccaggctca agctatcctt gcacctcagc ctcccaagta 3360

tgactaattt tttaaatatt tgagacgggg 3420

tctccctgtg ttacccaggc tggtctcaaa ctcctgggct caagtgatcc tcccatcttg 3480

gcctcccaga gtattgggat tacagacatg agccactgca cctgcccagc tccccaactc 3540

cctgccattt tttaagagac agtttcgctc catcgcccag gcctgggatg cagtgatgtg 3600

atcatagctc actgtaacct caaactctgg ggctcaagca gttctcccac cagcctcctt 3660

tttatttttt tgtacagatg gggtcttgct atgttgccca agctggtctt aaactcctgg 3720

cctcaagcaa tccttctgcc ttggcccccc aaagtgctgg gattgtgggc atgagctgct 3780

gtgcccagcc tccatgtttt aatatcaact ctcactcctg aattcagttg ctttgcccaa 3840

gataggagtt ctctgatgca gaaattattg ggctctttta gggtaagaag tttgtgtctt 3900

tgtctggcca catcttgact aggtattgtc tactctgaag acctttaatg gcttccctct 3960

ttcatctcct gagtatgtaa cttgcaatgg gcagctatcc agtgacttgt tctgagtaag 4020

tgtgttcatt aatgtttatt tagctctgaa gcaagagtga tatactccag gacttagaat 4080

agtgcctaaa gtgctgcagc caaagacaga gcggaactat gaaaagtggg cttggagatg 4140

gcaggagagc ttgtcattga gcctggcaat ttagcaaact gatgctgagg atgattgagg 4200

tgggtctacc tcatctctga aaattctgga aggaatggag gagtctcaac atgtgtttct 4260

gacacaagat ccgtggtttg tactcaaagc ccagaatccc caagtgcctg cttttgatga 4320

tgtctacaga aaatgctggc tgagctgaac acatttgccc aattccaggt gtgcacagaa 4380

aaccgagaat attcaaaatt ccaaattttt ttcttaggag caagaagaaa atgtggccct 4440

aaagggggtt agttgagggg tagggggtag tgaggatctt gatttggatc tctttttatt 4500

taaatgtgaa tttcaacttt tgacaatcaa agaaaagact tttgttgaaa tagctttact 4560

gtttctcaag tgttttggag aaaaaaatca accctgcaat cactttttgg aattgtcttg 4620

atttttcggc agttcaagct atatcgaata tagttctgtg tagagaatgt cactgtagtt 4680

ttgagtgtat acatgtgtgg gtgctgataa ttgtgtattt tctttggggg tggaaaagga 4740

4800

attttgttaa accat 4815

<210> 74

<211> 882

<212> PRT

<213> Homo sapiens

<400> 74

Met Gly Pro Trp Ser Arg Ser Leu Ser Ala Leu Leu Leu Leu Leu Gln

1 5 10 15

Val Ser Ser Trp Leu Cys Gln Glu Pro Glu Pro Cys His Pro Gly Phe

20 25 30

Asp Ala Glu Ser Tyr Thr Phe Thr Val Pro Arg Arg His Leu Glu Arg

35 40 45

Gly Arg Val Leu Gly Arg Val Asn Phe Glu Asp Cys Thr Gly Arg Gln

50 55 60

Arg Thr Ala Tyr Phe Ser Leu Asp Thr Arg Phe Lys Val Gly Thr Asp

65 70 75 80

Gly Val Ile Thr Val Lys Arg Pro Leu Arg Phe His Asn Pro Gln Ile

85 90 95

His Phe Leu Val Tyr Ala Trp Asp Ser Thr Tyr Arg Lys Phe Ser Thr

100 105 110

Lys Val Thr Leu Asn Thr Val Gly His His His Arg Pro Pro Pro His

115 120 125

Gln Ala Ser Val Ser Gly Ile Gln Ala Glu Leu Leu Thr Phe Pro Asn

130 135 140

Ser Ser Pro Gly Leu Arg Arg Gln Lys Arg Asp Trp Val Ile Pro Pro

145 150 155 160

Ile Ser Cys Pro Glu Asn Glu Lys Gly Pro Phe Pro Lys Asn Leu Val

165 170 175

Gln Ile Lys Ser Asn Lys Asp Lys Glu Gly Lys Val Phe Tyr Ser Ile

180 185 190

Thr Gly Gln Gly Ala Asp Thr Pro Val Gly Val Phe Ile Ile Glu

195 200 205

Arg Glu Thr Gly Trp Leu Lys Val Thr Glu Pro Leu Asp Arg Glu Arg

210 215 220

Ile Ala Thr Tyr Thr Leu Phe Ser His Ala Val Ser Ser Asn Gly Asn

225 230 235 240

Ala Val Glu Asp Pro Met Glu Ile Leu Ile Thr Val Thr Asp Gln Asn

245 250 255

Asp Asn Lys Pro Glu Phe Thr Gln Glu Val Phe Lys Gly Ser Val Met

260 265 270

Glu Gly Ala Leu Pro Gly Thr Ser Val Met Glu Val Thr Ala Thr Asp

275 280 285

Ala Asp Asp Asp Val Asn Thr Tyr Asn Ala Ala Ile Ala Tyr Thr Ile

290 295 300

Leu Ser Gln Asp Pro Glu Leu Pro Asp Lys Asn Met Phe Thr Ile Asn

305 310 315 320

Arg Asn Thr Gly Val Ile Ser Val Val Thr Thr Gly Leu Asp Arg Glu

325 330 335

Ser Phe Pro Thr Tyr Thr Leu Val Val Gln Ala Ala Asp Leu Gln Gly

340 345 350

Glu Gly Leu Ser Thr Thr Ala Thr Ala Val Ile Thr Val Thr Asp Thr

355 360 365

Asn Asp Asn Pro Pro Ile Phe Asn Pro Thr Thr Tyr Lys Gly Gln Val

370 375 380

Pro Glu Asn Glu Ala Asn Val Val Ile Thr Thr Leu Lys Val Thr Asp

385 390 395 400

Ala Asp Ala Pro Asn Thr Pro Ala Trp Glu Ala Val Tyr Thr Ile Leu

405 410 415

Asn Asp Asp Gly Gly Gln Phe Val Val Thr Thr Asn Pro Val Asn Asn

420 425 430

Asp Gly Ile Leu Lys Thr Ala Lys Gly Leu Asp Phe Glu Ala Lys Gln

435 440 445

Gln Tyr Ile Leu His Val Ala Val Thr Asn Val Val Pro Phe Glu Val

450 455 460

Ser Leu Thr Thr Ser Thr Ala Thr Val Thr Val Asp Val Leu Asp Val

465 470 475 480

Asn Glu Ala Pro Ile Phe Val Pro Pro Glu Lys Arg Val Glu Val Ser

485 490 495

Glu Asp Phe Gly Val Gly Gln Glu Ile Thr Ser Tyr Thr Ala Gln Glu

500 505 510

Pro Asp Thr Phe Met Glu Gln Lys Ile Thr Tyr Arg Ile Trp Arg Asp

515 520 525

Thr Ala Asn Trp Leu Glu Ile Asn Pro Asp Thr Gly Ala Ile Ser Thr

530 535 540

Arg Ala Glu Leu Asp Arg Glu Asp Phe Glu His Val Lys Asn Ser Thr

545 550 555 560

Tyr Thr Ala Leu Ile Ile Ala Thr Asp Asn Gly Ser Pro Val Ala Thr

565 570 575

Gly Thr Gly Thr Leu Leu Leu Ile Leu Ser Asp Val Asn Asp Asn Ala

580 585 590

Pro Ile Pro Glu Pro Arg Thr Ile Phe Phe Cys Glu Arg Asn Pro Lys

595 600 605

Pro Gln Val Ile Asn Ile Ile Asp Ala Asp Leu Pro Pro Asn Thr Ser

610 615 620

Pro Phe Thr Ala Glu Leu Thr His Gly Ala Ser Ala Asn Trp Thr Ile

625 630 635 640

Gln Tyr Asn Asp Pro Thr Gln Glu Ser Ile Ile Leu Lys Pro Lys Met

645 650 655

Ala Leu Glu Val Gly Asp Tyr Lys Ile Asn Leu Lys Leu Met Asp Asn

660 665 670

Gln Asn Lys Asp Gln Val Thr Thr Leu Glu Val Ser Val Cys Asp Cys

675 680 685

Glu Gly Ala Ala Gly Val Cys Arg Lys Ala Gln Pro Val Glu Ala Gly

690 695 700

Leu Gln Ile Pro Ala Ile Leu Gly Ile Leu Gly Gly Ile Leu Ala Leu

705 710 715 720

Leu Ile Leu Ile Leu Leu Leu Leu Leu Phe Leu Arg Arg Arg Ala Val

725 730 735

Val Lys Glu Pro Leu Leu Pro Pro Glu Asp Asp Thr Arg Asp Asn Val

740 745 750

Tyr Tyr Tyr Asp Glu Glu Gly Gly Gly Glu Glu Asp Gln Asp Phe Asp

755 760 765

Leu Ser Gln Leu His Arg Gly Leu Asp Ala Arg Pro Glu Val Thr Arg

770 775 780

Asn Asp Val Ala Pro Thr Leu Met Ser Val Pro Arg Tyr Leu Pro Arg

785 790 795 800

Pro Ala Asn Pro Asp Glu Ile Gly Asn Phe Ile Asp Glu Asn Leu Lys

805 810 815

Ala Ala Asp Thr Asp Pro Thr Ala Pro Pro Tyr Asp Ser Leu Leu Val

820 825 830

Phe Asp Tyr Glu Gly Ser Gly Ser Glu Ala Ala Ser Leu Ser Ser Leu

835 840 845

Asn Ser Ser Glu Ser Asp Lys Asp Gln Asp Tyr Asp Tyr Leu Asn Glu

850 855 860

Trp Gly Asn Arg Phe Lys Lys Leu Ala Asp Met Tyr Gly Gly Gly Glu

865 870 875 880

Asp Asp

<210> 75

<211> 3328

<212> DNA

<213> Homo sapiens

<400> 75

ccggaagtgc tgcgagccct gggccacgct ggccgtgctg gcagtgggcc gcctcgatcc 60

ctctgcagtc tttcccttga ggctccaaga ccagcaggtg aggcctcgcg gcgctgaaac 120

cgtgaggccc ggaccacagg ctccagatgg accctgggaa ggacaaagag ggggtgcccc 180

agccctcagg gccgccagca aggaagaaat ttgtgatacc cctcgacgag gatgaggtcc 240

ctcctggagt ggccaagccc ttattccgat ctacacagag ccttcccact gtggacacct 300

cggcccaggc ggcccctcag acctacgccg aatatgccat ctcacagcct ctggaagggg 360

ctggggccac gtgccccaca gggtcagagc ccctggcagg agagacgccc aaccaggccc 420

tgaaacccgg ggcaaaatcc aacagcatca ttgtgagccc tcggcagagg ggcaatcccg 480

tactgaagtt cgtgcgcaat gtgccctggg aatttggcga cgtaattccc gactatgtgc 540

tgggccagag cacctgtgcc ctgttcctca gcctccgcta ccacaacctg cacccagact 600

acatccatgg gcggctgcag agcctgggga agaacttcgc cttgcgggtc ctgcttgtcc 660

aggtggatgt gaaagatccc cagcaggccc tcaaggagct ggctaagatg tgtatcctgg 720

ccgactgcac attgatcctc gcctggagcc ccgaggaagc tgggcggtac ctggagacct 780

acaaggccta tgagcagaaa ccagcggacc tcctgatgga gaagctagag caggacttcg 840

tctcccggtc tctggaacag ctcatcgccg catcaagaga agatctggcc ttatgcccag 900

gcctgggccc tcagaaagcc cggaggctgt ttgatgtcct gcacgagccc ttcttgaaag 960

taccctgatg accccagctg ccaaggaaac ccccagtgta ataataaatc gtcctcccag 1020

gccaggctcc tgctggctgc gctggtgcag tctctgggga gggattctgg gggtgtcacc 1080

ttctggtggc ccaggtggggc accttcagct ttctttagtt cctcagtttc ccgggggcag 1140

actacacagg ctgctgctgc tgctgcttcc gcttcttgtc ccggcctgtg ggagcctcct 1200

ccccagactc tgaattcagt ggcggccctg gcatctcctc ttggggcact gtctctggca 1260

tccggctttc ctgactctgc ttcttcctct tcttggtgga tcccggagtt gccctggctt 1320

caggctgtcc ctcccctggc agttcaggct ctagtggctg aattggctca gtcactgtgt 1380

gacctctctc tttcttcttc ttcttcttct tggtggatgt gggagctgcc tgaggctcaa 1440

ggtcatccgg cagctcaggc cccaccacct ctgtctctgg ctccactgtg gcatcttgct 1500

1560

ctccttctgg ctgttgaggc gccatggtcc cccctgggga ctccagaggc ttcatctccg 1620

gctccactgg ctccatcgcc tccgtccctg gctccatcat tgccatctgt cccttttctt 1680

ttttcctctt cttcgtaggg ggcagaggga tggcttcctc cagtggctcc accttcacct 1740

gtggctgaga ctcaactgtc accccctcct ctggctccat cccttccgtc cccttttgcc 1800

1860

ccctgcttcac tgtcttgtct tctggctcga aggtttcttt ccctttggggc ttcttcctct 1920

tcttggtggt ggacgggaac agcactccca gaggctccag tgtctccact gtgggctctg 1980

tccccacagg ccctgctgcc tctggttctt tcagctgctg attttttttc ttcttcttct 2040

tccgcacatc catttctggc gaccccaaag ccatgtccac ctcggggcc ccgtgcccat 2100

2160 tcactgcctc ctgagtgact ggggcctctg tcacctgcat

aggtgagcag gttgggggcc aaggctgacc taggccctgt gactggtggg ttgcccccaa 2220

aggcacagaa ccgaggcctc aggccaggag ggatctgtgg tgggggactt gctgggatgg 2280

gctgcagagg gctccctgac agggattgct ggggaccctc aaggatcctt agggtgccct 2340

ggggggctga ggcacaggtg agtccacctc ctgcctccgt tgaggggggcc agcagggtcg 2400

cttctccagc ttggggacag ctgctgagga ctcgatagcg gtgccgcttg cctgccaatt 2460

2520

2580

tgtttttttt tgagagagtc ttgctctgtt gcctaggctg gagtgcagtg gcatgatctc 2640

ggctcactgc aatgtccacc tcctgggttc aagcgattct cctgcctcag cctctcaagt 2700

agctgggatt acaggcacct gccaccacgc ccagccaatt tttgtatttt tagtagagac 2760

aatttcacta tgttggccag gctggtcttg aactcctgac ctcaagtgat ctgctcgcct 2820

cggcctccca aggatggga ttacaggcac cagccactgc gcctggctgg cctctggttt 2880

ttaataaaac atgactagag tgactccatc ttaaagtgag tagctaggca cttacaaggt 2940

tcatgcttat ggcctgaaaa taaccacatc ccaggctgac caccaattat aattacagaa 3000

tatttatggc catacagaac atgttccacc aagcctgcag aatgtccaaa tgtcctaaga 3060

atgcagcccc cattacttaa atataacata aatgagcaag cttaggttgc aggattaatg 3120

gtcgtggata acaccaatag cccctacctt tagtgagctt atctgcacac tccaagttta 3180

actatagttc cttatagttt cttataagta gaaatactaa caaagggctg tgggtttctc 3240

cccctgcttt ctgaggacac tctactctgt aaaggagtag tttccaataa acttgtttct 3300

ttcactgtgc aaaaaaaaaaaaaaaaaa 3328

<210> 76

<211> 273

<212> PRT

<213> Homo sapiens

<400> 76

Met Asp Pro Gly Lys Asp Lys Glu Gly Val Pro Gln Pro Ser Gly Pro

1 5 10 15

Pro Ala Arg Lys Lys Phe Val Ile Pro Leu Asp Glu Asp Glu Val Pro

20 25 30

Pro Gly Val Ala Lys Pro Leu Phe Arg Ser Thr Gln Ser Leu Pro Thr

35 40 45

Val Asp Thr Ser Ala Gln Ala Ala Pro Gln Thr Tyr Ala Glu Tyr Ala

50 55 60

Ile Ser Gln Pro Leu Glu Gly Ala Gly Ala Thr Cys Pro Thr Gly Ser

65 70 75 80

Glu Pro Leu Ala Gly Glu Thr Pro Asn Gln Ala Leu Lys Pro Gly Ala

85 90 95

Lys Ser Asn Ser Ile Ile Val Ser Pro Arg Gln Arg Gly Asn Pro Val

100 105 110

Leu Lys Phe Val Arg Asn Val Pro Trp Glu Phe Gly Asp Val Ile Pro

115 120 125

Asp Tyr Val Leu Gly Gln Ser Thr Cys Ala Leu Phe Leu Ser Leu Arg

130 135 140

Tyr His Asn Leu His Pro Asp Tyr Ile His Gly Arg Leu Gln Ser Leu

145 150 155 160

Gly Lys Asn Phe Ala Leu Arg Val Leu Leu Val Gln Val Asp Val Lys

165 170 175

Asp Pro Gln Gln Ala Leu Lys Glu Leu Ala Lys Met Cys Ile Leu Ala

180 185 190

Asp Cys Thr Leu Ile Leu Ala Trp Ser Pro Glu Glu Ala Gly Arg Tyr

195 200 205

Leu Glu Thr Tyr Lys Ala Tyr Glu Gln Lys Pro Ala Asp Leu Leu Met

210 215 220

Glu Lys Leu Glu Gln Asp Phe Val Ser Arg Ser Leu Glu Gln Leu Ile

225 230 235 240

Ala Ala Ser Arg Glu Asp Leu Ala Leu Cys Pro Gly Leu Gly Pro Gln

245 250 255

Lys Ala Arg Arg Leu Phe Asp Val Leu His Glu Pro Phe Leu Lys Val

260 265 270

Pro

<210> 77

<211> 2204

<212> DNA

<213> Homo sapiens

<400> 77

ggagccccgg ctcctaggct gacagaccag cccagatcca gtggcccggga ggggcctgag 60

ctaaatccgc aggacctggg taacacgagg aagtcggttt ggtcccttta gggctccgga 120

tatctttggt gacttgtcca ctccagtgtg gcatcatgtg gcagctgctc ctcccaactg 180

ctctgctact tctagtttca gctggcatgc ggactgaaga tctcccaaag gctgtggtgt 240

tcctggagcc tcaatggtac agggtgctcg agaaggacag tgtgactctg aagtgccagg 300

gagcctactc ccctgaggac aattccacac agtggtttca caatgagagc ctcatctcaa 360

gccaggcctc gagctacttc attgacgctg ccacagtcga cgacagtgga gagtacaggt 420

gccagacaaa cctctccacc ctcagtgacc cggtgcagct agaagtccat atcggctggc 480

tgttgctcca ggcccctcgg tgggtgttca aggaggaaga ccctattcac ctgaggtgtc 540

acagctggaa gaacactgct ctgcataagg tcacatattt acagaatggc aaaggcagga 600

agtattttca tcataattct gacttctaca ttccaaaagc cacactcaaa gacagcggct 660

cctacttctg cagggggctt tttgggagta aaaatgtgtc ttcagagact gtgaacatca 720

ccatcactca aggtttggca gtgtcaacca tctcatcatt ctttccacct gggtaccaag 780

tctctttctg cttggtgatg gtactccttt ttgcagtgga cacaggacta tatttctctg 840

tgaagacaaa cattcgaagc tcaacaagag actggaagga ccataaattt aaatggagaa 900

aggaccctca agacaaatga cccccatccc atgggggtaa taagagcagt agcagcagca 960

tctctgaaca tttctctgga tttgcaaccc catcatcctc aggcctctct acaagcagca 1020

ggaaacatag aactcagagc cagatccctt atccaactct cgacttttcc ttggtctcca 1080

gtggaaggga aaagcccatg atcttcaagc agggaagccc cagtgagtag ctgcattcct 1140

agaaattgaa gtttcagagc tacacaaaca ctttttctgt cccaaccgtt ccctcacagc 1200

1260

taaattaccc agtttagagg ggaaaaaaaa acaattattc ctaaataaat ggataagtag 1320

aattaatggt tgaggcagga ccatacagag tgtgggaact gctggggatc tagggaattc 1380

agtgggacca atgaaagcat ggctgagaaa tagcaggtag tcgggatag tctaagggag 1440

gtgttcccat ctgagcccag agataagggt gtcttcctag aacattagcc gtagtggaat 1500

taacaggaaa tcatgagggt gacgtagaat tgagtcttcc aggggactct atcagaactg 1560

gaccatctcc aagtatataa cgatgagtcc tcttaatgct aggagtagaa aatggtccta 1620

ggaaggggac tgaggattgc ggtggggggt ggggtggaaa agaaagtaca gaacaaaccc 1680

tgtgtcactg tcccaagttg ctaagtgaac agaactatct cagcatcaga atgagaaagc 1740

ctgagaagaa agaaccaacc acaagcacac aggaaggaaa gcgcaggagg tgaaaatgct 1800

ttcttggcca gggtagtaag aattagaggt taatgcaggg actgtaaaac caccttttct 1860

gcttcaatat ctaattcctg tgtagctttg ttcattgcat ttattaaaca aatgttgtat 1920

aaccaatact aaatgtacta ctgagcttcg ctgagttaag ttatgaaact ttcaaatcct 1980

tcatcatgtc agttccaatg aggtggggat ggagaagaca attgttgctt atgaaagaaa 2040

gctttagctg tctctgtttt gtaagcttta agcgcaacat ttcttggttc caataaagca 2100

ttttacaaga tcttgcatgc tactcttaga tagaagatgg gaaaaccatg gtaataaaat 2160

2204

<210> 78

<211> 254

<212> PRT

<213> Homo sapiens

<400> 78

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Phe

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

Val Ser Phe Cys Leu Val Met Val Leu Leu Phe Ala Val Asp Thr Gly

210 215 220

Leu Tyr Phe Ser Val Lys Thr Asn Ile Arg Ser Ser Thr Arg Asp Trp

225 230 235 240

Lys Asp His Lys Phe Lys Trp Arg Lys Asp Pro Gln Asp Lys

245 250

<---

Claims (53)

1. Method for determining whether a cancer patient having a tumor antigen positive tumor is responding to treatment with an anti-tumor antigen antibody where the tumor antigen is a CLDN18.2 protein; said method comprising determining the genotype for one or more single nucleotide polymorphisms selected from the group consisting of MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260, and ERCC1 rs11615 in a sample obtained from a patient, characterized in that (a) the presence of a homozygous MUC1 rs4072037 [AA] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; (b) the presence of the homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk that the cancer patient is not a responder to antibody treatment; (c) the presence of a homozygous IL-10 rs1800896 [GG] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; (d) the presence of a heterozygous DNMT3A rs1550117 [GA] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; (e) the presence of a heterozygous SMAD4 rs12456284 [GA] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; (f) the presence of a homozygous EGF rs4444903 [AA] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; (g) the presence of a homozygous CDH1 rs16260 [AA] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment; and (h) the presence of a homozygous ERCC1 rs11615 [TT] genotype indicates a reduced risk that a cancer patient is not a responder to antibody treatment. 2. A method for determining whether there will be a progression-free survival period in a cancer patient having a tumor that is positive for a tumor antigen, where the tumor antigen is a CLDN18.2 protein; said method includes determining the genotype for one or more single nucleotide polymorphisms selected from the group consisting of MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260 and ERCC1 rs11615 in a sample obtained from a patient, where (a) the presence of a homozygous MUC1 rs4072037 [AA] genotype indicates a reduced risk that a patient with cancer will not have a progression-free survival period; (b) the presence of a homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk that a cancer patient will not have a progression-free survival period; (c) the presence of a homozygous IL-10 rs1800896 [GG] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period; (d) the presence of a heterozygous DNMT3A rs1550117 [GA] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period; (e) the presence of a heterozygous SMAD4 rs12456284 [GA] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period; (f) the presence of a homozygous EGF rs4444903 [AA] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period; (g) the presence of a homozygous CDH1 rs16260 [AA] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period; and (h) The presence of a homozygous ERCC1 rs11615 [TT] genotype indicates a reduced risk that a cancer patient will not have a progression-free survival period. 3. Method of determination (i) whether a cancer patient having a tumor antigen positive tumor responds to treatment with an anti-tumor antigen antibody, and (ii) whether there will be a progression-free survival period in a cancer patient who has a tumor antigen positive tumor, wherein the tumor antigen is a CLDN18.2 protein; said method includes determining the genotype for one or more single nucleotide polymorphisms selected from the group consisting of MUC1 rs4072037, IL-10 rs1800896, DNMT3A rs1550117, SMAD4 rs12456284, EGF rs4444903, CDH1 rs16260 and ERCC1 rs11615 in a sample obtained from a patient, where (a) the presence of the homozygous MUC1 rs4072037 [AA] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period; (b) the presence of a homozygous MUC1 rs4072037 [GG] genotype indicates an increased risk that the cancer patient is not a responder to antibody treatment and/or an increased risk that the cancer patient will not have a progression-free survival period; (c) the presence of a homozygous IL-10 rs1800896 [GG] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period; (d) the presence of a heterozygous DNMT3A rs1550117 [GA] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period; (e) the presence of a heterozygous SMAD4 rs12456284 [GA] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient will not have a progression-free survival period; (f) the presence of a homozygous EGF rs4444903 [AA] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient does not have a progression-free survival period; (g) the presence of a homozygous CDH1 rs16260 [AA] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient does not have a progression-free survival period; and (h) the presence of a homozygous ERCC1 rs11615 [TT] genotype indicates a reduced risk that the cancer patient is not a responder to antibody treatment and/or a reduced risk that the cancer patient does not have a progression-free survival period. 4. The method according to any one of paragraphs. 1-3, characterized in that the antibody acts by recruiting the patient's immune system to destroy tumor cells. 5. The method according to any one of paragraphs. 1-4, characterized in that the antibody acts through antibody-dependent cell-mediated cytotoxicity (ADCC) and/or complementary-dependent cytotoxicity (CDC). 6. The method according to any one of paragraphs. 1-5, characterized in that the antibody contains a heavy chain containing the amino acid sequence represented by SEQ ID NO: 17 or 51, or a fragment thereof, and a light chain containing the amino acid sequence represented by SEQ ID NO: 24, or its fragment. 7. A method of treating a cancer patient having an antigen positive tumor, wherein the tumor antigen is a CLDN18.2 protein, said method comprising a. evaluating whether a cancer patient is a responder to treatment with an antibody using the method of any one of claims. 1 and 3-6, and b. (i) treating the cancer patient with an antibody if the patient has a reduced risk of not responding to antibody treatment, or (ii) not treating the cancer patient with the antibody and/or treating the cancer patient with a treatment regimen that includes a treatment that is different from antibody treatment if the patient has an increased risk of not responding to antibody treatment. 8. The method according to claim 7, characterized in that the treatment regimen includes treatment that is independent of the patient's immune system. 9. The method of claim. 7 or 8, characterized in that the treatment regimen does not include treatment with an antibody that acts by recruiting the patient's immune system to destroy tumor cells. 10. The method according to any one of paragraphs. 7-9, characterized in that the treatment regimen includes surgery, chemotherapy and/or radiation. 11. The method according to any one of paragraphs. 7-10, characterized in that the treatment regimen includes treatment with a small molecular weight inhibitor of the tumor antigen and/or an antibody-drug conjugate, where the antibody is directed against the tumor antigen. 12. The method of claim 11, wherein the antibody-drug conjugate is an antibody linked to a radioactive, chemotherapeutic, or toxin component. 13. The method of claim 11 or 12, wherein the antibody-drug conjugate is an antibody bound to a cytostatic or cytotoxic compound. 14. The method according to any one of paragraphs. 1-13, characterized in that the tumor is a solid tumor. 15. The method according to any one of paragraphs. 1-14, characterized in that the tumor is a gastroesophageal tumor. 16. The method according to any one of paragraphs. 1-15, characterized in that the tumor is a progressive adenocarcinoma of the stomach or lower esophagus.

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