Tumor-specific Claudin 18.2 antibody

Abstract

The present invention provides a tumor-specific anti-CLDN18.2 antibody or a fragment thereof. The antibody or its fragment exhibits increased binding to tumor tissues expressing CLDN18.2 as compared to healthy tissues expressing CLDN18.2. In addition, the antibody does not exhibit cross-reactivity with CLDN18.1. The present invention also provides nucleic acids, vectors, host cells and medical uses.

Description

BACKGROUND OF THE INVENTION

[0002] Tight junctions are multiprotein complexes that connect adjacent epithelial or endothelial cells to form a barrier that prevents molecules from passing between cells and helps maintain the polarity of cells and tissues. Tight junctions are composed of three major groups of transmembrane proteins: claudins and occludins, cytoplasmic plaque proteins, and zonula occludens proteins. They also contain cytoskeletal proteins and signaling proteins such as actin, myosin II, and PKCζ. These proteins interact to maintain the tight junction structure (Yu and Turner 2008).

[0003] Claudins form a family of 23 proteins (Hewitt, Agarwal, and Morin 2006). Claudin 18 is a human protein encoded by the CLDN18 gene that forms tight junction strands in epithelial cells. Human CLDN18 can be alternatively spliced in the presence of two alternative first exons, resulting in two protein isoforms, CLDN18.1 (or claudin 18.1) and CLDN18.2 (or claudin 18.2). CLDN18.2 was first disclosed as the Zsig28 protein in WO2000 / 015659. The two isoforms differ in the N-terminal 69 amino acids that span the first extracellular loop. The first extracellular domain spans from amino acid 28 to amino acid 80. Within this range, there are 8 amino acid differences between CLDN18.1 and CLDN18.2. These two different isoforms are expressed in different tissues, with CLDN18.1 mainly expressed in lung tissue, while CLDN18.2 shows gastric specificity (Niimi et al. 2001). The expression of CLDN18.2 in normal stomach is limited to the short-lived differentiated cells of the gastric epithelium. The expression of CLDN18.2 has been further characterized in various tumor tissues. For example, CLDN18.2 has been found to be expressed in tumors of the pancreas, esophagus, ovary, and lung, associated with different histological subtypes (Sahin et al. 2008). The amino acid sequence of the human CLDN18.2 protein can be derived from the NCBI reference sequence: NP_001002026.1. This sequence is also disclosed as SEQ ID NO: 133.

[0004] Given the restricted expression pattern of CLDN18.2 in normal tissues and its ectopic expression in human cancers, it is an attractive cancer target for antibody therapy of epithelial tumors. Many studies have been conducted on this antibody therapy. WO2004 / 047863 identified splice variants of CLDN18 and screened antibodies against different peptides derived from CLDN18.2: peptide DQWSTQDLYN (SEQ ID NO:57), the N-terminal extracellular region of CLDN18.2, glycosylation-independent; peptide NNPVTAVFNYQ (SEQ ID NO:58), the N-terminal extracellular region of CLDN18.2, largely unglycosylated; and peptide STQDLYNNPVTAVF (SEQ ID NO:59), the N-terminal extracellular domain of CLDN18.2, unglycosylated. It also disclosed polyclonal rabbit antibodies screened with the pan-CLDN18 peptide TNFWMSTANMYTG (SEQ ID NO:60) in the C-terminal extracellular domain common to CLDN18.1 and CLDN18.2 isotypes. WO2005 / 113587 disclosed antibodies against specific epitopes of CLDN18.2 defined by peptide sequences: ALMIVGIVLGAIGLLV (SEQ ID NO:61) and RIGSMEDSAKANMTLTSGIMFIVS (SEQ ID NO:62). WO2007 / 059997 disclosed CLDN18.2-specific monoclonal antibodies obtained by immunization with the peptide METDTLLLWVLLLWVPGSTGDAAQPARRARRTKLGTELGSTPVWWNSADGRMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRAAIQHSGGRSRRARTKTHLRRGSE (SEQ ID NO:63), including the first extracellular domain of CLDN18.2 with N-terminal and C-terminal extensions. The antibodies obtained by this immunization mediate cell killing through complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC). The antibody IMAB362, also known as Claudiximab or Zolbetuximab, was disclosed in WO2007 / 059997 and WO2016 / 165762. IMAB362 is an IgG1 antibody derived from a murine monoclonal antibody and has been chimerized to display the human IgG1 constant region for clinical use.WO2008 / 145338 also discloses antibodies that bind to overlapping peptides (MDQWSTQDLYNNPVT (SEQ ID NO:64), LYNNPVTAVFNYQGL (SEQ ID NO:65), VFNYQGLWRSCVRES (SEQ ID NO:66), QGLWRSCVRESSGFT (SEQ ID NO:67), and RSCVRESSGFTECRG (SEQ ID NO:68)) within the first extracellular domain. For the diagnostic purpose of generating antibodies that target the C-terminal portion of CLDN18.2 for detecting CLDN18.2 expression in cells of cancer tissue sections, WO2013 / 167259 discloses antibodies that bind to C-terminal epitopes of CLDN18.2. The sequences of these two epitopes are TEDEVQSYPSKHDYV (SEQ ID NO:69) and EVQSYPSKHDYV (SEQ ID NO:70). WO2013 / 174509 proposes combinations of anti-CLDN18.2 antibodies with agents that stabilize γδ T cells or with agents that stabilize or increase CLDN18.2 expression. The antibodies can be conjugated to therapeutic moieties such as cytotoxins, drugs (e.g., immunosuppressants), or radioisotopes. WO2014 / 075788 discloses methods of treating cancer diseases using bispecific antibodies that bind CLDN18.2 and CD3. WO2014 / 127906 discloses combinatorial agents that stabilize or increase CLDN18.2 expression. WO2016 / 166122 discloses anti-CLDN18.2 monoclonal antibodies that can be efficiently internalized after CLDN18.2 binding and are thus suitable for antibody-drug conjugate (ADC) development. In addition, the conjugation of such antibodies to the drugs DM4 and MMAE using cleavable SPDB or valine-citrulline linkers is also disclosed. However, despite the disclosure of all the antibodies in the patent applications, currently only the chimeric IMAB362 disclosed in WO2007 / 059997 and WO2016 / 165762 has been tested in clinical trials. In addition to these antibodies and ADCs, WO2018 / 006882 also discloses chimeric antigen receptors (CARs) based on anti-CLDN18.2 monoclonal antibodies. The antibodies of WO2018 / 006882 have been humanized and their sequences are disclosed in the supplementary material section related to Jiang et al. 2018 (Jiang et al. 2018). CAR T cells based on the humanized antibodies are currently being tested in a phase I clinical trial (ClinicalTrials.gov identifier: NCT03159819) in patients with advanced gastric adenocarcinoma and pancreatic cancer. CN109762067 discloses other anti-CLDN18.2 monoclonal antibodies that mediate cell killing via CDC and ADCC.WO2019 / 173420 discloses a humanized monoclonal antibody against CLDN18.2 with ADCC activity. WO2019 / 175617 discloses a monoclonal antibody against CLDN18.2 that binds to a different epitope compared to IMAB362. WO2019 / 219089 discloses a monoclonal antibody that binds to a CLDN18.2 mutant.

[0005] CLDN18.2 has been described to exist in different conformations and contain potential extracellular N-glycosylation sites (see WO2007 / 059997, page 3, first paragraph), which may lead to potentially different topologies / differential glycosylation between normal and tumor cells (see WO2007 / 059997, page 4, second paragraph). However, none of the reported antibodies preferentially target CLDN18.2 expressed on tumor cells. Since CLDN18.2 is expressed not only in tumors but also in healthy tissues, namely in gastric tissue (Sahin et al. 2008), it is clearly beneficial to have an antibody that only targets CLDN18.2 expressed in tumors to avoid the safety issues and side effects often associated with the off-target effects of therapeutic antibodies on healthy organs / tissues (Hansel et al. 2010), especially as reported for IMAB362 (Sahin et al. 2018; Tureci et al. 2019).

[0006] In addition to binding to the target with high affinity, therapeutic antibodies should also maintain their desired properties during development, production, storage, and clinical application (in vivo). Post-translational modifications (PTMs) may impair the stability of antibodies (Lu et al. 2019; Gervais 2016). Since uncontrolled PTMs may result in the efficacy, activity, potency, or stability of the antibody being lower than desired, it is very important to design antibodies with the minimum possible PTMs during the development of therapeutic antibodies. PTMs can also have a profound impact on the regulatory acceptance, technology transfer, or processes and development of biosimilars. The main modifications are oxidation, deamidation, and isomerization. In addition, IMAB362 is a chimeric antibody that still has extended mouse sequences, which may lead to the production of anti-drug antibodies in some patients, which may, for example, result in a decrease in therapeutic efficacy after repeated applications.

[0007] Therefore, there is a need for improved antibodies specific for CLDN18.2 for the treatment of cancer patients.

[0008] Definitions

[0009] "Antibody", also known as "immunoglobulin" (Ig), typically comprises four polypeptide chains - two heavy (H) chains and two light (L) chains, and is thus a multimeric protein, or comprises its equivalent Ig homologs (e.g., camelid antibodies that contain only heavy chains, single domain antibodies (sdAbs) or nanobodies, which can be derived from heavy or light chains). The term "antibody" includes antibody-based binding proteins, modified antibody forms that retain target binding ability. The term "antibody" also includes full-length functional mutants, variants or derivatives (including but not limited to murine, chimeric, humanized and fully human antibodies) that retain the basic epitope binding characteristics of the Ig molecule, and includes bispecific, bivalent, multispecific and dual variable domain Igs. The Ig molecule can be of any class (e.g., IgG, IgE, IgM, IgD, IgA and IgY) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) and allotype. The Ig molecule can also be mutated, e.g., to enhance or reduce affinity for Fcγ receptors or the neonatal Fc receptor (FcRn).

[0010] As used herein, "antibody fragment" refers to a non-full-length molecule that contains at least one polypeptide chain derived from an antibody and exhibits target binding. Antibody fragments are capable of binding the same epitope or target as their corresponding full-length antibody. Antibody fragments include, but are not limited to, (i) Fab fragments, which are monovalent fragments consisting of a variable light (VL) domain, a variable heavy (VH) domain, a constant light (CL) domain and a constant heavy 1 (CH1) domain, alone or in any combination; (ii) F(ab')2 fragments, which are bivalent fragments containing two Fab fragments linked by a disulfide bond in the hinge region (reduction of an F(ab')2 fragment yields two Fab' fragments with free sulfhydryl groups); (iii) the heavy chain portion of a Fab(Fa) fragment, which consists of the VH and CH1 domains; (iv) variable fragment (Fv) fragments, which consist of the VL and VH domains of a single arm of an antibody; (v) domain antibody (dAb) fragments, which contain a single variable domain; (vi) isolated complementarity determining regions (CDRs); (vii) single-chain Fv fragments (scFv); (viii) diabodies, which are bivalent bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain but the linker used is too short to allow pairing between the two domains on the same chain, thus forcing the domains to pair with complementary domains on the other chain and generating two antigen binding sites; (ix) linear antibodies, which contain a pair of tandem Fv fragments (VH-CH1-VH-CH1) that together with a complementary light chain polypeptide form a pair of antigen binding regions; (x) dual variable domain immunoglobulins; (xi) other non-full-length portions of immunoglobulin heavy and / or light chains, or mutants, variants or derivatives thereof.

[0011] As used herein, "antibody-based binding protein" can represent any protein that contains at least one antibody-derived VH, VL, or CH immunoglobulin domain in the context of other non-immunoglobulin or non-antibody-derived components. Such antibody-based proteins include, but are not limited to, (i) Fc fusion proteins of binding proteins, including receptors or receptor components having all or part of an immunoglobulin CH domain, (ii) binding proteins in which the VH and / or VL domains are coupled to alternative molecular scaffolds, or (iii) molecules in which the immunoglobulin VH and / or VL and / or CH domains are combined and / or assembled in a manner not normally found in native antibodies or antibody fragments.

[0012] As used herein, the term "modified antibody forms" encompasses antibody-drug conjugates (ADCs), polyalkylene oxide-modified scFvs, monobodies, diabodies, camelid antibodies, domain antibodies, bispecific or trispecific antibodies, IgA, or two IgG structures linked by a J chain and a secretory component, shark antibodies, New World primate frameworks and non-New World primate CDRs, IgG4 antibodies with the hinge region removed, IgG with two additional binding sites engineered into the CH3 domain, antibodies with an altered Fc region to enhance or reduce affinity for Fcγ receptors, dimeric constructs containing CH3, VL, and VH, and the like.

[0013] The Kabat numbering scheme (Martin and Alleman 2014) has been applied to the disclosed antibodies.

[0014] When the term "comprising" is used in the present specification and claims, it does not exclude other elements. For the purposes of the present invention, the term "consisting of" is considered a preferred embodiment of the term "comprising". If a group is defined hereinafter as including at least a certain number of embodiments, this should also be understood as disclosing a group consisting preferably only of these embodiments.

[0015] Unless otherwise expressly stated, the indefinite or definite articles used in connection with a singular noun, such as "a", "an", or "the", include the plural forms of that noun.

[0016] Technical terms are used in their common sense. If a particular meaning is to be conveyed to certain terms, the definition of the term will be given in the context in which the term is used hereinafter. DETAILED DESCRIPTION OF THE INVENTION

[0018] The inventors have surprisingly identified novel anti-CLDN18.2 antibodies as further described in the following embodiments, which exhibit increased binding to tumor cells expressing CLDN18.2 compared to healthy gastric cells expressing CLDN18.2 and / or have improved stability and / or are humanized while retaining their improved properties.

[0019] Thus, in one embodiment of the present invention, the present invention provides an antibody or a fragment thereof that binds to CLDN18.2, wherein the antibody or the fragment thereof exhibits increased binding to tumor tissue expressing CLDN18.2 as compared to healthy tissue expressing CLDN18.2. In one embodiment, the healthy cells or tissues for comparison are healthy gastric cells or healthy gastric tissue.

[0020] As shown in Examples 4 and 5 respectively, the increased binding of the antibody or the fragment thereof provided herein to tumor tissue can be demonstrated by bioanalytical methods such as flow cytometry (FC) or immunohistochemistry (IHC). Tumors expressing CLDN18.2 can be generated by subcutaneous injection of A549 cells expressing CLDN18.2 into Balb / c mice. A549 cells expressing CLDN18.2 can be generated as shown in Example 4 and can be obtained under the accession number DSM ACC3360 at DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, 38124 Braunschweig, DE, deposited on December 6, 2019. Healthy tissues (such as healthy gastric tissue) can also be obtained from tumor - bearing mice. Thus, the increased binding to tumor tissue as compared to healthy tissue can be demonstrated on tumor tissue and healthy tissue obtained from the same animal.

[0021] The increased binding to CLDN18.2 expressed in tumor tissue as compared to CLDN18.2 expressed in healthy tissue may be due to post - translational modifications such as differential glycosylation of CLDN18.2 or misfolding of CLDN18.2.

[0022] Flow cytometry (FC) can be used as a bioanalytical method for testing antibody binding. The percentage of CLDN18.2 - positive cells can be measured, for example, by FC with a specific anti - CLDN18.2 antibody. Another possible binding readout can be, for example, the ratio of the percentage of CLDN18.2 - positive cells in a tumor cell sample to the percentage of CLDN18.2 - positive cells in a cell sample obtained from healthy tissue (such as healthy gastric tissue). The increased binding of the antibody to CLDN18.2 - expressing tumor cells generated from A549 cells expressing CLDN18.2 as compared to healthy cells (such as healthy gastric cells) can be demonstrated by a ratio of >2, >5, ≥10, preferably ≥15, and more preferably ≥20.

[0023] Compared with healthy cells (such as healthy gastric cells), the binding of the antibody to tumor cells expressing CLDN18.2 generated from A549 cells expressing CLDN18.2 is increased, and can also be described by showing that the antibody binds at least 2-fold more, at least 5-fold more, at least 10-fold more, preferably at least 15-fold more, preferably at least 20-fold more tumor cells compared to healthy cells (such as healthy gastric cells).

[0024] Immunohistochemistry (IHC) can be used as a bioanalytical method to test antibody binding. Preferably, tissue samples for IHC should be snap-frozen after resection and, once thawed, fixed in acetone as shown, for example, in Example 5. Since CLDN18.2 is a tight junction protein in healthy tissue, positive CLDN18.2 staining should result in an appearance dominated by membrane staining at the cell-cell interface in healthy tissue and / or tumor tissue. Thus, negative or weak CLDN18.2 staining should result in a lack of membrane staining.

[0025] In another embodiment, the present invention provides an antibody or a fragment thereof that binds CLDN18.2 with a half-maximal effective concentration (EC50) value higher than 0.4 μg / ml, higher than 0.5 μg / ml, preferably higher than 0.6 μg / ml, but not higher than 1 μg / ml when measured by flow cytometry (FC) titration of HEK293T cells overexpressing CLDN18.2. HEK293T cells overexpressing CLDN18.2 can be generated as described in Example 3. When measured by flow cytometry (FC) titration of HEK293T cells overexpressing CLDN18.2, the EC50 value of the antibody of the present invention can be 0.4 to 1 μg / ml, 0.5 to 1 μg / ml, or preferably 0.6 to 1 μg / ml.

[0026] Alternatively, when measured by flow cytometry of HEK293T cells overexpressing CLDN18.2, the EC50 value of the antibody of the present invention can be compared with the EC50 value of IMAB362, where the EC50 value of the antibody of the present invention is at least 1.1-fold higher, at least 1.2-fold higher, preferably at least 1.5-fold higher, more preferably at least 2-fold higher, even more preferably at least 2.5-fold higher than the EC50 value of IMAB362, but not more than 5-fold higher than the EC50 value of IMAB362. When measured by flow cytometry of HEK293T cells overexpressing CLDN18.2, the EC50 value of the antibody of the present invention can be 1.1-fold to 2.5-fold higher, 1.2-fold to 2.5-fold higher, preferably 1.5-fold to 2.5-fold higher, or more preferably 2-fold to 2.5-fold higher than the EC50 value of IMAB362.

[0027] In another embodiment, the present invention provides an antibody or fragment thereof that binds CLDN18.2 with an EC50 value of greater than 0.6 μg / ml, greater than 1 μg / ml, preferably greater than 1.5 μg / ml, more preferably greater than 2 μg / ml but not greater than 3 mg / ml when measured by flow cytometry titration of PA-TU-8988S-High cells. PA-TU-8988S-High cells can be generated as described in Example 2. When measured by flow cytometry titration of PA-TU-8988S-High cells, the EC50 value of the antibody of the present invention can be 0.6 to 3 μg / ml, 1 to 3 mg / ml, preferably 1.5 to 3 μg / ml, or more preferably 2 to 3 μg / ml.

[0028] Alternatively, when measured by flow cytometry of PA-TU-8988S-High cells, the EC50 value of the antibody of the present invention can be compared to the EC50 value of IMAB362, wherein the EC50 value of the antibody of the present invention is at least 1.5-fold higher, at least 2-fold higher, preferably at least 3-fold higher, more preferably at least 4-fold higher than the EC50 value of IMAB362, but not more than 5-fold higher. When measured by flow cytometry of PA-TU-8988S-High cells, the EC50 value of the antibody of the present invention can be 1.5-fold to 5-fold higher, 2-fold to 5-fold higher, 3-fold to 5-fold higher, or 4-fold to 5-fold higher than the EC50 value of IMAB362.

[0029] In another embodiment, the present invention provides an antibody or fragment thereof that binds CLDN18.2 with a maxMFI value within + / - 40% of the maxMFI value of IMAB362 when measured by flow cytometry of HEK293T cells overexpressing CLDN18.2. The present invention also provides an antibody or fragment thereof that binds CLDN18.2 with a maxMFI value equal to or up to 2-fold higher than the maxMFI value of IMAB362 when measured by flow cytometry of PA-TU-8988S-High cells.

[0030] Antibodies or functional fragments thereof that exhibit increased binding to tumor tissues expressing CLDN18.2 compared to healthy tissues expressing CLDN18.2 may have a therapeutic advantage over antibodies that cannot distinguish between healthy tissues and tumor tissues expressing CLDN18.2. Tumor-specific antibodies may not cause safety issues and side effects typically associated with on-target effects of therapeutic antibodies in healthy organs / tissues (Hansel et al. 2010). For example, for IMAB362, such unwanted effects have been reported (Sahin et al. 2018; Tureci et al. 2019).

[0031] The present invention also provides an antibody or fragment thereof that binds to CLDN18.2, which comprises heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:21, SEQ ID NO:22, and SEQ ID NO:23, respectively, and light chain CDRs LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, respectively.

[0032] The present invention also provides an antibody or fragment thereof that binds to CLDN18.2, which comprises the heavy chain HCDR3 sequence of SEQ ID NO:23 and the light chain LCDR3 sequence of SEQ ID NO:26.

[0033] The corresponding consensus sequences can be found in Table 1. It should be understood that any antibody or fragment thereof that binds to CLDN18.2 based on any combination of CDRs derived from the consensus sequences is part of the present invention.

[0034] Table 1: Isolated antibody CDR consensus sequences

[0035]

[0036]

[0037] Antibody binding or binding affinity is typically expressed by the equilibrium binding or dissociation constant (K a or K d ), which are in turn the reciprocal ratio of the dissociation and association rate constants (k off and k on ), respectively. Thus, equivalent affinities can correspond to different rate constants as long as the ratio of the rate constants remains constant. Binding affinity and / or rate constants can be determined using techniques well known in the art or described herein, such as ELISA, flow cytometry titration, isothermal titration calorimetry (ITC), Biacore (SPR), biolayer inferometry, or fluorescence polarization. In some cases, due to the nature of the antigen, it may be difficult to measure the K a or K d of an antibody. This is especially true for integral membrane proteins such as claudins (Hashimoto et al. 2018). In such cases, the integral membrane protein can be expressed as proteoliposomes or lipid particles. Such lipid particles can be immobilized on plastic and used in ELISA assays to determine the binding affinity of the antibody to the immobilized antigen. Thus, the half-maximal effective concentration (EC50) value of each test antibody or its functional fragment can be calculated instead of Ka or K d value, reflecting its binding affinity (or binding strength) to the antigen. Example 2 and Figure 1 below illustrate the ELISA assay binding affinity curves of antibodies having CDRs contained in the consensus sequences of Table 1. The EC50 value and the maximum binding value can be used to quantify the binding of the antibody to CLDN18.2. Example 3 below relates to calculating the EC50 value of an antibody having CDRs contained in the consensus sequences of Table 1 by flow cytometry of cells expressing CLDN18.2.

[0038] In another embodiment, the present invention provides an antibody or fragment thereof that binds to CLDN18.2 and comprises heavy chain CDR HCDR1, HCDR2, and HCR3 sequences of SEQ ID NO:21, SEQ ID NO:126, and SEQ ID NO:23, respectively, and light chain CDR LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, respectively.

[0039] In one embodiment, the present invention relates to an antibody or fragment thereof that binds to CLDN18.2 and comprises:

[0040] a. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:15, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively;

[0041] b. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively;

[0042] c. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:17, SEQ ID NO:14, and SEQ ID NO:11, respectively;

[0043] d. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:16 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:11 respectively;

[0044] e. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:15 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively;

[0045] f. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively;

[0046] g. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:11 respectively;

[0047] h. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively; or

[0048] i. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:17, SEQ ID NO:14 and SEQ ID NO:11 respectively.

[0049] In yet another embodiment, the present invention provides an antibody or fragment thereof that binds to CLDN18.2 and comprises:

[0050] a. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively;

[0051] b. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:7, and SEQ ID NO:8 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11 respectively; or

[0052] c. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:2, and SEQ ID NO:3 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:11 respectively.

[0053] In yet another embodiment, the present invention relates to an antibody or fragment thereof that binds to CLDN18.2 and comprises:

[0054] a. The VH sequence of SEQ ID NO:27 and the VL sequence of SEQ ID NO:28;

[0055] b. The VH sequence of SEQ ID NO:29 and the VL sequence of SEQ ID NO:30;

[0056] c. The VH sequence of SEQ ID NO:31 and the VL sequence of SEQ ID NO:32.

[0057] In another embodiment, the present invention relates to an antibody or fragment thereof that binds to CLDN18.2 and comprises:

[0058] a. The VH sequence of SEQ ID NO:33;

[0059] b. The VH sequence of SEQ ID NO:34;

[0060] c. The VH sequence of SEQ ID NO:35;

[0061] d. The VH sequence of SEQ ID NO:36; or

[0062] e. The VH sequence of SEQ ID NO:37;

[0063] and

[0064] f. The VL sequence of SEQ ID NO: 38;

[0065] g. The VL sequence of SEQ ID NO: 39;

[0066] h. The VL sequence of SEQ ID NO: 40; or

[0067] i. The VL sequence of SEQ ID NO: 41.

[0068] In another embodiment, the present invention relates to an antibody or a fragment thereof that binds to CLDN18.2 and comprises:

[0069] a. The VH sequence of SEQ ID NO: 33 and the VL sequence of SEQ ID NO: 38;

[0070] b. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 38;

[0071] c. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 39;

[0072] d. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 40;

[0073] e. The VH sequence of SEQ ID NO: 35 and the VL sequence of SEQ ID NO: 38;

[0074] f. The VH sequence of SEQ ID NO: 36 and the VL sequence of SEQ ID NO: 41;

[0075] g. The VH sequence of SEQ ID NO: 36 and the VL sequence of SEQ ID NO: 40;

[0076] h. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 41;

[0077] i. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 38; or

[0078] j. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 39.

[0079] In another embodiment, the present invention relates to an antibody that binds to CLDN18.2 and comprises:

[0080] a. The heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51;

[0081] b. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:51;

[0082] c. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:52;

[0083] d. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:53;

[0084] e. The heavy chain sequence of SEQ ID NO:48 and the light chain sequence of SEQ ID NO:51;

[0085] f. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:54;

[0086] g. The heavy chain sequence of SEQ ID NO:49 and the light chain sequence of SEQ ID NO:53;

[0087] h. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:54;

[0088] i. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:51; or

[0089] j. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:52.

[0090] The constant light chain region CL, the constant heavy chain region CH1 and the Fc region of the disclosed antibody may respectively have the amino acid sequences of SEQ ID NO:127 and SEQ ID NO:128.

[0091] In a preferred embodiment, the present invention relates to an antibody that binds to CLDN18.2 and comprises the heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51.

[0092] In a further preferred embodiment, the present invention relates to an antibody that binds to CLDN18.2 and consists of the heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51.

[0093] The present invention also relates to antibodies having an amino acid sequence with at least 80% identity, at least 85% identity, at least 90% identity, at least 95% identity, or at least 98% identity to the amino acid sequence of the antibody of the present invention, which exhibit increased binding to tumor cells expressing CLDN18.2 as compared to healthy gastric cells expressing CLDN18.2.

[0094] In one embodiment, the present invention relates to an antibody that binds to CLDN18.2 and has an amino acid sequence with at least 80% identity, at least 85% identity, at least 90% identity, at least 95% identity, or at least 98% identity to an antibody comprising:

[0095] a. The VH sequence of SEQ ID NO:27 and the VL sequence of SEQ ID NO:28;

[0096] b. The VH sequence of SEQ ID NO:29 and the VL sequence of SEQ ID NO:30;

[0097] c. The VH sequence of SEQ ID NO:31 and the VL sequence of SEQ ID NO:32.

[0098] In a further embodiment, the present invention relates to an antibody that binds to CLDN18.2 and has an amino acid sequence with at least 80% identity, at least 85% identity, at least 90% identity, at least 95% identity, or at least 98% identity to an antibody comprising:

[0099] a. The VH sequence of SEQ ID NO:33 and the VL sequence of SEQ ID NO:38;

[0100] b. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:38;

[0101] c. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:39;

[0102] d. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:40;

[0103] e. The VH sequence of SEQ ID NO:35 and the VL sequence of SEQ ID NO:38;

[0104] f. The VH sequence of SEQ ID NO:36 and the VL sequence of SEQ ID NO:41;

[0105] g. The VH sequence of SEQ ID NO:36 and the VL sequence of SEQ ID NO:40;

[0106] h. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:41;

[0107] i. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:38; or

[0108] j. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:39.

[0109] In yet another embodiment, the invention relates to an antibody that binds to CLDN18.2 and has an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 98% identical to the antibody consisting of the heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51.

[0110] In another embodiment, the Fc domain of the antibody (or the antibody fragment present) can contain modifications or mutations such as those listed in Table 2 below. Such modifications or mutations can be introduced to modulate the effector activity of the Fc domain of the antibody. Modifications of the antibody can also include peptide tags added to the C-terminus of the antibody HC chain and / or LC chain. Such tags can be used, for example, for protein purification or protein conjugation.

[0111] In another embodiment, the present invention provides an anti-CLDN18.2 antibody or fragment thereof, which antibody is IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4, synthetic IgG, IgM, F(ab)2, Fv, scFv, IgGACH2, F(ab')2, scFvCH3, Fab, VL, VH, scFv4, scFv3, scFv2, dsFv, Fv, scFv-Fc, (scFv)2, non-depleting IgG, diabody, bivalent antibody or an Fc-engineered version thereof. In a preferred embodiment, the antibody is an antibody of the IgG1 type. The Fc region of immunoglobulins interacts with various Fcγ receptors (FcγR) and complement proteins (e.g., C1q) and mediates immune effector functions, such as elimination of target cells by antibody-dependent cell cytotoxicity (ADCC), antibody-dependent cell phagocytosis (ADCP) or complement-dependent cytotoxicity (CDC). For therapeutic methods, enhancing or silencing the relevant effector functions may be beneficial. The type of immunoglobulin (IgA, IgD, IgE, IgG, IgM) can be selected according to the desired effector function of the antibody associated with the Fc domain. Synthetic immunoglobulins can also be used, such as immunoglobulins having IgG2 amino acids 118 to 260 and IgG4 amino acids 261 to 447 or IgG2 variants having point mutations from IgG4 (e.g., H268Q / V309L / A30S / P331S). Such synthetic immunoglobulins reduce the effector function of the antibody. Fc-engineered immunoglobulins can also be used to modulate antibody effector functions. Table 2 shows examples of such Fc engineering. Expression in production cell lines with altered fucosylation can also affect FcγR binding.

[0112] Table 2 : Examples of modifications that modulate antibody effector functions. Unless otherwise noted, mutations are on the IgG1 subclass (Wang, Mathieu and Brezski 2018).

[0113]

[0114]

[0115] The half-life of an antibody can also be adjusted. The Fc domain plays a central role in the stability and serum half-life of an antibody. For therapeutic applications, the antibody half-life can be shortened by using antibody fragments that lack the Fc domain or have a truncated Fc domain, such as F(ab)2, Fv, scFv, IgGACH2, F(ab')2, scFvCH3, Fab, VL, VH, scFv4, scFv3, scFv2, dsFv, Fv, scFv-Fc or (scFv)2. The antibody can also be in the form of a diabody or a bivalent antibody. Diabodies or bivalent antibodies can be used to increase the affinity for the target, thereby allowing for lower doses. Functional fragments that lack the Fc domain or have a truncated Fc domain can also be used to develop other therapeutic methods, such as chimeric antigen receptor T cells (CAR T cells) or bispecific T cell engagers (BiTE). In a CAR construct, one VH domain and one VL domain are typically linked by a short peptide linker to form a single-chain variable fragment (scFv), and the scFv fragment is further linked to a transmembrane domain and other domains of the cytoplasmic T cell immunoreceptor tyrosine-based activation motif (from, for example, CD3ζ) and co-stimulatory molecules (from, for example, CD28, 4-1BB (CD127) or OX40) (Chang and Chen 2017). The VH and VL domains used in the scFv fragment can be the antibodies listed in Table 3. A BiTE typically consists of a fusion of two scFvs from two different antibodies. One scFv domain can be an isolated antibody that binds CLDN18.2 listed in Table 3, while the other scFv domain is from an antibody that binds, for example, CD3, CD16, NKG2D, NKp46, CD2, CD28 or CD25. Adequate guidance on BiTE antibody formats and other bispecific antibody formats for T cell redirection can be found in the review by Diego Ellerman (2019).

[0116] In another embodiment, the present invention provides an antibody or a fragment thereof that binds CLDN18.2, the antibody having a constant light chain region (CL) of SEQ ID NO: 127 and preferably a constant heavy chain region CH1 of SEQ ID NO: 129 with reduced FcγR binding and an Fc region having an L234A / L235A mutation in the constant heavy chain region CH2. More preferably, the present invention provides an antibody having a constant heavy chain region CH1 and an Fc region of SEQ ID NO: 130, which has an L234A / L235A / P329G mutation in the constant heavy chain region CH1 and the Fc region, with even further reduced FcγR binding.

[0117] In another preferred embodiment, the invention relates to an antibody or fragment thereof that binds to CLDN18.2 and comprises the VH sequence of SEQ ID NO:33, the VL sequence of SEQ ID NO:38, the constant light chain region (CL) of SEQ ID NO:127, and the constant heavy chain region CH1 and Fc region of SEQ ID NO:129 having L234A / L235A.

[0118] In another preferred embodiment, the invention relates to an antibody or fragment thereof that binds to CLDN18.2 and consists of the VH sequence of SEQID NO:33, the VL sequence of SEQ ID NO:38, the constant light chain region (CL) of SEQ ID NO:127, and the constant heavy chain region CH1 and Fc region of SEQ ID NO:129 having L234A / L235A.

[0119] In another embodiment, the invention provides an antibody or fragment thereof that binds to CLDN18.2, wherein the antibody or fragment thereof is humanized. The humanization of monoclonal antibodies is well established. The Handbook of TherapeuticAntibodies, 2nd edition provides sufficient information on the humanization of monoclonal antibodies (Saldanha 2014), bioinformatics tools for analyzing such antibodies (Martin and Alleman 2014), and the development and preparation of therapeutic antibodies (Jacobi et al. 2014).

[0120] In another embodiment, the antibody or fragment thereof is an isolated antibody or isolated fragment that binds to CLDN18.2.

[0121] In another embodiment, the invention provides an antibody or fragment thereof that binds to CLDN18.2, wherein the antibody or fragment thereof does not bind to CLDN18.1. Thus, the antibody does not exhibit cross-reactivity or cross-binding with CLDN18.1. The binding of the antibody to the target protein can be tested by flow cytometry of cells expressing the target protein. The specific binding of the tested antibody to its target protein can be visualized on a histogram. This graph produces a peak with a high fluorescence signal when the antibody specifically binds to the expressed target protein, while it produces a peak with a low fluorescence signal when the antibody does not bind or binds only very weakly to the expressed target protein. The degree of binding can also be represented by a bar graph showing the maximum mean fluorescence intensity (maxMFI) measured by flow cytometry, where a high maxMFI reflects strong binding and a low / no maxMFI reflects no binding or very weak binding. Comparing the maxMFI values of different antibodies in the same experimental setup can also indicate the affinity of the antibody for the target, with a higher maxMFI indicating a lower off rate and higher affinity. In Example 3 and Figure 4and Figure 5 Examples of this binding assay can be found therein.

[0122] In another embodiment, the present invention provides an antibody or fragment thereof that binds to CLDN18.2, which antibody binds to another moiety. The binding of the antibody or fragment thereof to the other moiety can be covalent or non-covalent. The moiety can include a radioisotope, a fluorescent tag, a histological marker, a cytotoxin, or a cytokine. The covalent binding of the moiety to the antibody can be facilitated by a linker known in the art.

[0123] In yet another embodiment, the present invention relates to a tumor-specific antibody or fragment thereof that binds to CLDN18.2, wherein the antibody is less susceptible to post-translational deamidation than IMAB362. In another embodiment, the present invention relates to a tumor-specific antibody or fragment thereof that binds to CLDN18.2, wherein the antibody does not undergo post-translational deamidation. Post-translational modifications (PTMs) are important issues in antibody development as well as antibody production and storage. Uncontrolled PTMs can result in antibodies with lower efficacy, activity, potency, or stability. PTMs can be N-glycosylation, lysine glycosylation, and cysteine capped with other cysteine, glutathione, or other thiol-containing compounds from the cell culture medium during bioprocessing, or the formation of dimers and higher-order oligomers due to cysteines linked by covalent disulfide bonds. Among PTMs, deamidation of asparagine (Asn, N) residues, isomerization of aspartate (aspartic acid, Asp, D) residues, and the formation of succinimide intermediates are the most common modification reactions of therapeutic antibodies in vivo during production, storage, or after administration. Deamidation of Asn and isomerization of Asp depend on sequence propensity, structural environment, and storage conditions, especially solution pH and storage temperature. These modifications can lead to a decrease or even loss of function or biological activity, especially when the affected residues are involved in target binding. Asn and Asp residues are at risk of modification, especially when they are located in regions of structural flexibility (such as CDR loops) and meet certain other structural prerequisites, while the framework regions have been observed to have comparable resistance to modification. In addition to the structural positions of Asn and Asp residues, typical motifs for Asn deamidation and Asp isomerization have been identified. These typical motifs are NG, NS, NN, NT, NH, and DG, DS, DD, DT, and DH (Lu et al. 2019), respectively. In a computational analysis, the disclosed antibodies present the DG Asp isomerization motif in the last amino acid of CDR2 in the VL domain and in the CH2 and CH3 regions of HC (VL-CDR2 (at position 62), CH2 (at position 282), CH3 (at position 403)).

[0124] The isomerization of Asp can be tested by placing the antibody at low pH (i.e., pH 5.5) and heating (i.e., 40 °C) for two weeks, while the deamidation of Asn in the antibody can be tested by placing the antibody at high pH (i.e., pH 8.0) and heating (i.e., 40 °C) for one week to simulate the production and storage conditions.

[0125] The inventors have now shown that, despite these harsh conditions, the disclosed antibodies contain Asn and Asp in their CDRs and carry an Asp-Gly (DG) Asp-isomerization motif, but surprisingly they do not have Asn deamidation (see Table 6) and Asp isomerization (see Table 7) and their binding affinity to CLDN18.2 is not affected. On the other hand, IMAB362 shows Asn deamidation under these conditions, inducing a loss of binding affinity (as shown in Table 6 and Figure 10 as shown). Accordingly, the present invention provides an isolated antibody or fragment thereof that binds to CLDN18.2 and is less susceptible to PTM compared to IMAB362 during production, storage, and clinical application (in vivo), and ensures the maintained binding affinity to CLDN18.2 during production, storage, and clinical application (in vivo).

[0126] The present invention also provides an antibody that binds to the same epitope as the antibodies described herein. In one embodiment, the antibody binds to the same epitope as the antibody comprising the heavy chain sequence of SEQ ID NO: 46 and the light chain sequence of SEQ ID NO: 51.

[0127] The present invention further provides an antibody that competes with the antibodies described herein for binding. In one embodiment, the antibody competes for binding with the antibody comprising the heavy chain sequence of SEQ ID NO: 46 and the light chain sequence of SEQ ID NO: 51.

[0128] The present invention further provides an antibody that competitively inhibits the binding of the antibodies described herein to Claudin 18.2. In one embodiment, the antibody competitively inhibits the binding of the antibody comprising the heavy chain sequence of SEQ ID NO: 46 and the light chain sequence of SEQ ID NO: 51 to Claudin 18.2.

[0129] Suitable methods for detecting the binding of an antibody to the same antigen include methods for localizing antigen-antibody interactions. This method has been described in Abbott 2014 (Abbott, Damschroder, and Lowe 2014). Suitable methods for detecting competition include competitive assays by epitope binning, as described in Abdiche 2009 (Abdiche et al. 2009). Suitable methods for detecting competitive inhibition include ELISA assays.

[0130] According to one embodiment, the present invention provides nucleic acid sequences encoding isolated tumor-specific antibodies or functional fragments thereof that bind to CLDN18.2. The nucleic acid sequences can encode individual CDRs, encode VH and VL regions, or encode the entire heavy and light chains of the antibody. These nucleic acid sequences can be found in Table 3. The nucleic acid sequences can also encode F(ab)2, Fv, scFv, IgGACH2, F(ab')2, scFvCH3, Fab, VL, VH, scFv4, scFv3, scFv2, dsFv, Fv, scFv-Fc, (scFv)2, non-depleting IgG, diabody, bivalent antibody, or Fc-engineered versions thereof. The encoded immunoglobulin can be IgA1, IgA2, IgD, IgE, IgG1, IdG2, IgG3, IgG4, synthetic IgG, IgM, or mutant and Fc-engineered versions thereof.

[0131] In yet another embodiment, the nucleic acid sequences can also encode a CAR construct that binds to CLDN18.2. Adequate guidance for constructing CAR T cells can be found in Chang and Chen (2017) or June and Sadelain (2018). In one embodiment, the present invention provides T cells that are genetically engineered to produce an artificial T cell receptor, such as a chimeric antigen receptor (CAR), wherein the artificial T cell receptor comprises an antibody or a functional fragment thereof that binds to CLDN18.2 of the present invention.

[0132] In yet another embodiment, the present invention provides tumor-specific antibody-based binding proteins that specifically bind to CLDN18.2. Such binding proteins can comprise at least the CLDN18.2-binding domain of the disclosed antibody and another protein domain that is unrelated to the antibody. The present invention also provides modified antibody forms that bind to CLDN18.2.

[0133] The present invention also provides expression vectors that comprise the nucleic acids of the present invention or degenerate nucleic acids resulting from codon degeneracy. The expression vectors can be expression vectors for expressing proteins in mammalian cells, bacteria, fungi, or insect cells, and the host cell type for the expression vector carrying the nucleic acid encoding the antibody or its functional fragment is selected. Adequate guidance for constructing such vectors can be found in Green and Sambrook (Green and Sambrook 2012).

[0134] In another embodiment, the present invention provides host cells comprising the nucleic acids or expression vectors of the present invention. The host cells can be mammalian cells or cell lines, bacterial cells, fungal cells, or insect cells.

[0135] In another embodiment, the present invention relates to an antibody or fragment thereof that binds to CLDN18.2, a nucleic acid encoding the antibody or fragment thereof, a vector containing the nucleic acid, or a host cell containing the nucleic acid or the vector, for treating a subject suffering from a tumor disease.

[0136] In another embodiment, the present invention relates to an antibody or fragment thereof that binds to CLDN18.2, a nucleic acid encoding the antibody or fragment thereof, a vector containing the nucleic acid, or a host cell containing the nucleic acid or the vector, for treating a subject at risk of developing a neoplastic disease, and / or for treating a subject diagnosed with a neoplastic disease.

[0137] The disclosed antibody or fragment thereof can be used as a monotherapy. In a preferred embodiment, the disclosed antibody or fragment thereof is used in combination with the established standard of care for tumor diseases.

[0138] The tumor disease can be at least one disease selected from pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, and lung cancer. It is understood that the neoplastic disease to be treated expresses CLDN18.2.

[0139] In one embodiment, the subject is a mammal. In a preferred embodiment, the subject is a human.

[0140] Another embodiment of the present invention provides a method for treating a neoplastic disease using an antibody or functional fragment thereof that binds to CLDN18.2, the neoplastic disease including pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, or lung cancer, wherein the method comprises administering to a subject in need a pharmaceutically effective amount of the antibody or functional fragment thereof. The treatment method can be a monotherapy or preferably a combination therapy with the established standard of care for tumor diseases.

[0141] The amino acid sequence of the human CLDN18.2 protein can be derived from the NCBI reference sequence: NP_001002026.1. This sequence is also disclosed as SEQ ID NO:133. BRIEF DESCRIPTION OF THE DRAWINGS

[0142] Figure 1: Binding of selected chimeric and humanized anti-CLDN18.2 antibodies, as designated, to lipid particles containing CLDN18.2 or empty lipid particles was evaluated by ELISA. A. Chimeric antibodies cCl1-1, cCl1-2, cCl1-3, IMAB362, and secondary antibody only; B. Humanized antibodies hCl1a to hCl1j, chimeric cCl1-1, IMAB362, and secondary antibody only. All newly generated antibodies bind to liposomal CLDN18.2.

[0143] Figure 2 ：PA-TU-8988S cells were sorted according to the CLDN18.2 expression level. A. FC spectrum of PA-TU-9888S stained with IMAB362. B. FC spectrum of PA-TU-8988S cells sorted by FACS for high expression of CLDN18.2.

[0144] Figure 3 ： HEK293T cells overexpressing huCLDN18.2 were generated. HEK293T cells that do not endogenously express CLDN18.2 were transfected with plasmids encoding huCLDN18.2 for stable expression of CLDN18.2 or encoding huCLDN18.1 for stable expression of CLDN18.1. Expression was analyzed by FC after staining with IMAB362 and a pan-CLDN18.1 antibody or only an anti-human IgG secondary antibody. A. FC spectrum of untransfected HEK293T cells. B. FC spectrum of transfected HEK293T cells stably expressing CLDN18.1. C. FC spectrum of transfected HEK293T cells stably expressing CLDN18.2.

[0145] Figure 4 ： Flow cytometry binding assays of chimeric cCl1-1, cCl1-2, and cCl1-3 antibodies with pre-B cell L11 cells overexpressing CLDN18.1 or CLDN18.2. The chimeric antibodies bind to CLDN18.2 but not to CLDN18.1. IMAB362 was used as a positive binding control.

[0146] Figure 5 ： Flow cytometry binding assays of humanized hCl1a to hCl1j antibodies with HEK293T cells overexpressing CLDN18.1 or CLDN18.2. The humanized antibodies bind to CLDN18.2 but not to CLDN18.1. IMAB362 and cCL1-1 were used as positive binding controls.

[0147] Figure 6 ： FACS expression spectrum of A549 cells overexpressing CLDN18.2. A549 cells that do not express endogenous CLDN18.2 were stably transfected with a plasmid encoding CLDN18.2, and the expression of CLDN18.2 was analyzed by FACS using IMAB362.

[0148] Figure 7 ：Flow cytometry live cell staining. The figure represents the percentage of isolated single cells bound by CLDN18.2 antibodies (cCl1-1, hCl1a, hCl1b, hCl1c, hCl1f, and IMAB362). Single cells were isolated from mouse tumors expressing CLDN18.2 induced by injected A549 cells overexpressing CLDN18.2 (solid bars) or from healthy mouse stomachs expressing CLDN18.2 (hollow bars).

[0149] Figure 8 ： Frozen gastric tissue staining. Frozen tissue sections of healthy mouse gastric tissue expressing CLDN18.2 were stained with hCl1a (A), hCl1b (B), hCl1c (C), hCl1f (D), or IMAB362 (E) antibodies. The pictures are representative IHC images.

[0150] Figure 9 ： Staining of frozen tumor tissue induced by injected A549 cells overexpressing CLDN18.2. Frozen tissue sections of mouse tumors expressing CLDN18.2 were stained with hCl1a (A), hCl1f (B), IMAB362 (C), or Abcam34H14L15 pan-CLDN18 antibody. The pictures are representative IHC images.

[0151] Figure 10 ： Effect of deamidation on the binding activity of IMAB362. The affinity of IMAB362 for CLDN18.2 decreased after deamidation. Example

[0152] Example 1: Generation of Chimeric and Humanized Antibodies

[0153] Techniques for generating monoclonal antibodies are well established. The Handbook of Therapeutic Antibodies, 2nd Edition (2014) provides sufficient information on these techniques, such as generating monoclonal antibodies by immunizing mice or rats (Moldenhauer 2014), humanization of monoclonal antibodies (Saldanha 2014), bioinformatics tools for analyzing antibodies (Martin and Alleman 2014), or the development and preparation of therapeutic antibodies (Jacobi et al. 2014). Briefly, monoclonal antibodies against CLDN18.2 were generated by DNA immunization of rats with a plasmid encoding human CLDN18.2 cDNA (huCLDN18.2) (NCBI reference sequence: NM_001002026.3). The specific reactivity of rat immune sera against huCLDN18.2 was analyzed by flow cytometry (FC analysis) and ELISA. Hybridoma clones were then generated from lymphocytes isolated from immunized rats to obtain chimeric antibodies. Three clones were identified as CLDN18.2-specific, resulting in chimeric antibodies named cCl1-1, cCl1-2, and cCl1-3 with similar CDRs (see Table 3). Subsequently, cCl1-1, cCl1-2, and cCl1-3 were humanized, yielding 10 humanized clones named hCl1a, hCl1b0, hCl1c, hCl1d, hCl1e, hCl1f, hCl1g, hCl1h, hCl1i, and hCl1j antibodies (see Table 3).

[0154] As a control, the IMAB362 antibody was synthesized using the sequences of the heavy chain (SEQ ID NO:55) and light chain (SEQ ID NO:56) published in WO2013 / 174509, named monoclonal antibody 182-D1106-362, deposit number DSM ACC2810, deposited on October 26, 2006 at DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Inhoffenstr.7B 38124 Braunschweig DE.

[0155] Table 3: Antibody nucleic acid and amino acid sequences

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169] The antibodies described in Examples 2 to 5 are further modified to contain the RLPQTGG tag (SEQ ID NO: 131) at the C-terminus of the HC and / or the GGGGS LPQTGG tag (SEQ ID NO: 132) at the C-terminus of the LC. In this case, the C-terminal lysine (K) on the HC is replaced by the Arg (R) of the tag. The addition of the tag does not change the affinity and specificity of the antibody for CLDN18.2.

[0170] Example 2: ELISA Assays and FC Titrations to Confirm Binding of Chimeric and Humanized Antibody Variants to CLDN18.2

[0171] The binding affinities of the chimeric antibody and the humanized antibody (hCl) for CLDN18.2 were tested in an ELISA assay in which lipid particles carrying CLDN18.2 were used as the antigen source. CLDN18.2-lipid particles and empty lipid particles (without bound antigen, as a negative control) were used to coat a 96-well plate at a final concentration of 10 μg / ml. After washing with PBS / 0.05% Tween-20 (PBS-T) and blocking with PBS-T / 3% BSA at 37°C for at least 1 hour, 1:3 serial dilutions of the test antibody starting at a concentration of 2 μg / ml were added to the coated wells and incubated at 37°C for at least 1 hour. By binding an HRP-goat anti-human secondary antibody, in SIGMAFAST TMOPD was used as a peroxidase substrate for development. The reaction was terminated by adding 2M H2SO4, and then the OD at 490 nm was read on an ELISA plate reader, revealing the presence of the bound antibody. Representative binding curves are shown in Figure 1. All test antibodies of the present invention specifically bind to lipid particles containing CLDN18.2. Interestingly, humanization of the chimeric antibodies did not result in a decrease in affinity compared to the parental chimeric cCl1-1 antibody, as could be expected, and for 6 out of 10 antibodies, even increased their affinity.

[0172] Binding of chimeric and humanized antibodies to CLDN18.2 was also tested by FC titration using PA-TU-8988S cells (Creative Bioarray, catalog number CSC-C0326) overexpressing CLDN18.2 and HEK293T (ATCC, CRL-3216 TM ) cells. FC titration allows measurement of the half-maximal effective concentration (EC50) of the test antibody. PA-TU-8988S cells expressing high levels of CLDN18.2 were selected by FACS. In this article, these cells are named PA-TU-8988S-High cells. Based on FACS staining of IMAB362, the PA-TU-8988S cell population expresses different levels of CLDN18.2, with high-level and medium-level expression (see Figure 2 A). To have a more homogeneous cell population, the cells were sorted by FACS to select only cells with higher CLDN18.2 expression. Briefly, PA-TU-8988S cells suspended in FACS buffer (PBS, 2% FCS) were incubated with 2 μg / ml of IMAB362 on ice for 30 minutes. After washing in FACS buffer, the cells were incubated with a PE-labeled Fcγ-specific IgG goat anti-human secondary antibody (eBioscience) on ice for 30 minutes. After washing, the stained cells were resuspended in FACS buffer and analyzed and sorted by a FACSAria TM instrument to separate medium-expressing cells from high-expressing cells ( Figure 2 B). After sorting, the collected PA-TU-8988S-High cells were resuspended in growth medium, amplified, and frozen aliquots were stored in liquid nitrogen. HEK293T cells overexpressing CLDN18.2 or CLDN18.1 were generated as described in Example 3, and the expression of CLDN18.2 was analyzed by flow cytometry ( Figure 3 ).

[0173] To quantify the binding of the antibody to CLDN18.2, 250x10 3Overexpressing CLDN18.2 HEK293T cells or PA-TU-8988-High cells at [[number of cells per well]] cells / well were seeded into FC buffer (PBS / 2% FBS) in a 96-well plate and pelleted by centrifugation. IMAB362 and the hCl antibody to be tested were diluted at 20 μg / ml, then serially diluted 1:4 and incubated with the allotted cells for 30 minutes at 4 °C. After washing with FC buffer, a PE-conjugated anti-human IgG secondary antibody was added to the cells and incubated for an additional 30 minutes at 4 °C, then further washed with FC buffer. The cells were then resuspended in 100 μl of FC buffer and measured using a FACSCalibur TM cell analyzer (BD Biosciences, USA). FC analysis (see Figure 5 and Table 4) showed that the hCl antibody had a higher EC50 value than IMAB362, although its maxMFI value was in the same range as IMAB362. Similar maxMFI values can indicate similar binding / dissociation rates for IMAB362 and the hCl antibody.

[0174] Table 4: Maximum MFI and EC50 (μg / ml) of All hCl and IMAB362 Antibodies Measured on HEK293T Cell Lines and PA-TU-8988S-High Cell Lines Overexpressing CLDN18.2 Example 3: Generation of Pre-B Cell L11 Cells and HEK293 T Cells Stably Expressing hCLDN18.1 and hCLDN18.2; Testing of the Binding Specificity of Chimeric and Humanized Antibodies

[0175]

[0176] Figure 3 Figure 4

[0177] The pre-B cell line L11 (Waldmeier et al. 2016) and the HEK293T (ATCC CRL-3216TM) cell line do not endogenously express CLDN18.1 or CLDN18.2. Therefore, to test antibody binding, CLDN18.1 and CLDN18.2 were recombinantly overexpressed in these cell lines. Cells were co-transfected by electroporation with a transposase expression construct (pcDNA3.1-hy-mPB), constructs with transposable full-length huCLDN18.1 (pPB-Puro-huCLDN18.1) or huCLDN18.2 (pPB-Puro-huCLDN18.2) and a puromycin resistance cassette, and a construct carrying EGFP as a transfection control (pEGFP-N3) (Waldmeier et al. 2016). After electroporation, the cells were allowed to recover for two days in growth medium in a humidified incubator at 37 °C, in a 7.5% CO2 atmosphere for L11 cells and in a 5% CO2 atmosphere for HEK293T cells. Transfection was verified by FC analysis of EGFP expression. Cells expressing CLDN18.1 or CLDN18.2 were then selected by adding 1 μg / ml puromycin to the culture and further amplified to allow the generation of frozen stocks in FCS containing 10% DMSO. Expression of CLDN18.1 and CLDN18.2 in the transfected cells was analyzed by FC (see Figure 5 ). Briefly, trypsinized HEK293T cells and L11 cells growing in suspension were collected by centrifugation, resuspended in PBS / 2% FCS, and stained for 30 minutes on ice with IMAB362 as the primary antibody at a concentration of 2 μg / ml for CLDN18.2, then washed in PBS / 2% FCS and stained for 30 minutes on ice with anti-human IgG (Fcγ-specific) PE goat antibody (eBioscience) as the secondary antibody. After further washing, the stained cells resuspended in ice-cold FC buffer were analyzed using a FACSCalibur TM instrument (see Figure 3 and Figure 4 ). Un-transfected parental cells, which do not express CLDN18.2, were used as negative controls. Expression of CLDN18.1 was analyzed in a similar manner using a proprietary pan-CLDN18 antibody that recognizes CLDN18.1 and CLDN18.2 (see Figure 5) Any pan-CLDN18 antibody that can be used for flow cytometry measurement is also applicable, such as the antibody anti-Claudin-18 / CLDN18 (C-terminal) (Catalog No. AP50944PU-N) provided by OriGene Technologies, the CLDN18 (C-terminal) rabbit pAb (Catalog No. MBS8555451) from MyBioSource, or the CLDN18 antibody (Catalog No. 63-847) from ProSci.

[0178] Therefore, L11 and HEK293T cells stably expressing huCLDN18.1 and huCLDN18.2 were used to test the binding specificity of the chimeric antibodies cCl1-1, cCl1-2, cCl1-3 and the humanized antibody to CLDN18.2 rather than CLDN18.1. The cells were stained with 2 μg / ml of the antibody on ice for 30 minutes, then washed in PBS / 2% FCS and stained with anti-human IgG (Fcγ-specific) PE goat antibody (eBioscience) as the secondary antibody on ice for 30 minutes. All three chimeric antibodies ( Figure 5 ) and the humanized antibody ( Example 4: Testing of the Binding Activity of Humanized CLDN18.2 Antibodies by Flow Cytometry on Fresh Tumor Tissue and Fresh Gastric Tissue ) bound to huCLDN18.2 expressed by L11 or HEK293T cells, but not to huCLDN18.1. In addition, the affinity of the humanized antibody for binding to huCLDN18.2 was similar to that of IMAB362, and the affinity was at least as good as that of cCl1-1 ( Figure 6 ).

[0179] Figure 7 Table 5:

[0180] A549 (ATCC CCL-185 TM) The cell line does not endogenously express CLDN18.1 or CLDN18.2. To test the binding of the antibody to CLDN18.2, CLDN18.2 was expressed in A549 cells. A549 cells were co-transfected by electroporation with a transposase expression construct (pcDNA3.1-hy-mPB) (Klose et al. 2017), a construct with a transposable full-length huCldn18.2 (pPB-Puro-huCldn18.1) and a puromycin expression cassette, and a construct carrying EGFP as a transfection control (pEGFP-N3) (Waldmeier et al. 2016). After electroporation, the cells were allowed to recover for two days in the growth medium in a humidified incubator at 37 °C and 5% CO2 atmosphere. Transfection was verified by FC analysis of EGFP expression. Cells expressing CLDN18.1 or CLDN18.2 were then selected by adding 1 μg / ml puromycin to the culture and further amplified to allow the generation of frozen stocks in FCS containing 10% DMSO. Expression of CLDN18.2 in the transfected cells was analyzed by FC. Briefly, trypsinized A549 cells were collected by centrifugation, resuspended in PBS / 2% FCS and stained for 30 minutes on ice with IMAB362 as the primary antibody at a concentration of 2 μg / ml against CLDN18.2, then after washing in PBS / 2% FCS, stained for 30 minutes on ice with 2.5 μg / ml anti-human IgG (Fcγ specific) PE goat antibody (eBioscience) as the secondary antibody. After further washing, the stained cells resuspended in ice-cold FC buffer were analyzed using a FACSCalibur TM instrument (see Example 5: Testing of Humanized CLDN18.2 Antibodies by Immunohistochemistry (IHC) on Frozen Tissue Samples ). Un-transfected parental cells that do not express CLDN18.2 were used as negative controls. These cells were deposited on December 6, 2019 at DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Inhoffenstr. 7B, 38124 Braunschweig, DE and are available under the deposit number DSM ACC3360.

[0181] 1x10 6 A549 cells expressing CLDN18.2 in 100 μl of 50% Matrigel were implanted subcutaneously into two Balb / c mice, and tumor growth was monitored over several weeks until the tumors reached 150 - 450 mm 3The expected size between. Healthy gastric tissue and tumor tissue were collected for FC analysis. The collected tissues were cut into small pieces and digested with the Miltenyi Tumor Dissociation Kit (MACS Miltenyi Biotec, Germany). The tissue pieces were incubated with dissociation buffer (prepared according to the manufacturer's instructions) in a 6-well plate at 37 °C with continuous gentle shaking for 30 minutes. The samples were resuspended and filtered through a 70 μm cell strainer (Corning, USA), and then washed with 20 ml of FC buffer (PBS + 2% FBS). The cell suspension was centrifuged (400 g for 5 minutes at 4 °C) and the supernatant was discarded. If necessary, the cell suspension was filtered through a strainer and centrifuged repeatedly. The pellet was resuspended in 5 ml of red blood cell lysis buffer (Biolegend, USA) and incubated on ice for 4 minutes. After incubation, 25 ml of PBS was added and the suspension was centrifuged again (400 g for 5 minutes at 4 °C). The pellet was resuspended in FC buffer (0.5 - 3 ml based on the pellet). An equal number of cells were transferred to a 96-well plate and further processed for FC analysis. The cells in the plate were washed with PBS and centrifuged (400 g for 2 minutes at 4 °C). The pellet was resuspended in 50 μl / well of a staining mixture consisting of the selected antibodies diluted in PBS (4 μg / ml of cCl1-1, hCl1a, hCl1b, hCl1c, and hCl1f; 2 μg / ml of IMAB364) and an AF488-labeled AE1 / AE3 pan-cytokeratin antibody (Thermo Fisher Scientific, USA), and incubated on ice for 25 minutes. After incubation, the cells were washed twice in PBS and centrifuged (400 g for 2 minutes at 4 °C). The pellet was resuspended in 50 μl / well of a secondary staining mixture (PBS + PE-labeled anti-human antibody) (Thermo Fisher Scientific, USA) and incubated on ice for 25 minutes. After incubation, the cells were washed twice again in PBS. The pellet was resuspended in 100 μl of PBS containing DAPI. The plate was kept on ice until FC analysis. For FC analysis, live cells were separated from dead cells by forward scatter and DAPI staining. Then the live cells were gated to determine the presence of cytokeratin (AF888 positive) and bound CLDN18.2 antibody (PE positive cells). The results of the FC analysis can be seen in Figure 8 and Table 5. The results are the average of data obtained from two mice.

[0182] All tested antibodies (cCl1-1, hCl1a, hCl1b, hCl1c, hCl1f, and IMAB364) bind to a similar percentage (approximately between 20% and 30%) of tumor cells bearing CLDN18.2. However, surprisingly, only IMAB362 binds to healthy gastric cells bearing CLDN18.2, while the binding of cCl1-1, hCl1a, hCl1b, hCl1c, and hCl1f is barely detectable, binding to less than 1% of healthy gastric cells. The difference in binding ability between tumor cells derived from injected A549 cells expressing CLDN18.2 and CLDN18.2 expressed in healthy gastric cells is also expressed as the ratio of the percentage of positive tumor cells divided by the percentage of positive gastric cells (see the last column in Table 5). For IMAB362, this ratio is less than 5 and, on average, close to 1, while for the tested cCl1-1 humanized clones (hCl1a, hCl1b, hCl1c, and hCl1f), this ratio is higher than 15 and, on average, higher than 30.

[0183] Figure 9 FC binding data and binding ratios of selected antibodies to healthy gastric cells and tumor cells.

[0184]

[0185]

[0186] Thus, compared to healthy gastric cells, cCl1-1 and the tested cCl1-1 humanized clones (hCl1a, hCl1b, hCl1c, and hCl1f) show increased binding to tumor cells and are thus tumor-specific CLDN18.2 antibodies. In contrast, IMAB362 cannot distinguish between tumor cells bearing CLDN18.2 and healthy gastric cells bearing CLDN18.2.

[0187] Figure 8

[0188] From subcutaneous implantation of 1x10 6Fresh gastric and tumor tissue samples expressing CLDN18.2 obtained from Balb / c mice with A549 cells expressing CLDN18.2 were snap-frozen in OCT in a suitable tissue mold. Tissue sections 5 - 15 μm thick were cut with a cryostat at -20 °C, transferred to microscope slides at room temperature (RT), and then kept frozen until IHC staining. Before staining, the slides were returned to room temperature and fixed in pre-cooled acetone (-20 °C) for 10 minutes. After evaporation of acetone at room temperature, the slides were rinsed in TBS and processed to block non-specific staining sites: the slides were incubated in 0.3% H2O2 at room temperature for 15 minutes, then washed with TBS and incubated in a peroxidase blocking solution (Agilent, USA) at room temperature for 60 minutes. After blocking, the slides were subjected to antibody staining: the slides were incubated with primary antibodies (hCL1a, hCl1b, hCl1c, hCl1f, IMAB362, and 34H14L15 pan-CLDN18 antibody (Abcam, USA)) at room temperature for 120 minutes, washed in TBS, and then incubated with an HRP-conjugated anti-human antibody (or anti-rabbit antibody for the pan-CLDN18 antibody) at room temperature for 30 minutes. Antibodies bound to CLDN18.2 or pan-CLDN18 on the tissue sections were revealed by treating the slides with a DAB+ substrate Chromogen system (Agilent, USA) according to the manufacturer's instructions. After subsequent washing in TBS, the slides were counterstained with hematoxylin, rinsed in dH2O for 15 minutes, dehydrated in sequential 95% and 100% ethanol washes, and then the slides were further cleared in xylene. Finally, the slides were mounted with a coverslip in a glycerol mounting medium (Agilent, USA). Representative microscopic images of healthy mouse gastric tissue and mouse tumor tissue staining can be found separately in Figure 9 and Example 6: Analysis of the Asn Deamidation and Asp Isomerization Propensities of Humanized Antibody (hCl) Variants and IMAB362 respectively.

[0189] Table 6: shows representative staining of healthy gastric tissue. In tissues co-stained with hCL1a, hCl1b, hCl1c, and hCl1f (Figures A, B, C, and D respectively), only hematoxylin staining of the cell nuclei was visible, while tissues co-stained with IMAB362 (Figure E) showed membrane CLDN18.2 DAB staining. Thus, compared to IMAB362 binding to healthy gastric tissue expressing CLDN18.2, the humanized clones of cCl1-1 tested (hCL1a, hCl1b, hCl1c, and hCl1f) did not bind to healthy gastric tissue expressing CLDN18.2. In addition, Figure 10Representative staining of tumor tissues is shown. Panels A, B, C, and D are representative images of tumor tissues stained with hCl1a, hCl1f, IMAB362, and Abcam 34H14L15 pan-CLDN18 antibody, respectively. All tumors stained with the test antibodies showed strong membranous CLDN18.2 DAB staining. Similar to IMAB362 or the pan-CLDN18 antibody, the humanized clones of cCl1-1 (hCL1a and hCl1f) tested bound to mouse tumor tissues expressing CLDN18.2. Thus, compared to healthy gastric tissues expressing CLDN18.2, the humanized clones of cCl1-1 showed increased binding to tumor tissues expressing CLDN18.2.

[0190] Table 7:

[0191] Deamidation of Asn (N) residues and isomerization of Asp (D) residues can occur during biopharmaceutical preparation, storage, or clinical use (in vivo). Deamidation and isomerization can lead to potential changes in protein structure, function, activity, stability, and immunogenicity. Therefore, they must be minimized and controlled, especially in a regulatory environment. The presence of Asn deamidation and Asp isomerization motifs can be analyzed on a computer. The most common Asn deamidation motif is the NG motif, while the most common Asp isomerization motif is the DG motif.

[0192] This computational analysis revealed that all hCl antibodies possess a potential DG Asp isomerization motif in the second CDR of VL, and neither hCl antibodies nor IMAB362 possess a potential NG deamidation motif in their CDRs. To validate the computational predictions, hCl antibodies and IMAB362 were stressed under high or low pH and thermal conditions to accelerate modifications that may occur during the manufacturing process and long-term storage. Briefly, the antibody sample buffer was exchanged to 20 mM sodium phosphate buffer pH 8.0 for Asn deamidation stress testing or 20 mM citrate buffer, pH 5.5 for Asp isomerization stress testing using Amicon centrifugal filters, and the samples were diluted to a final concentration of 3.0 mg / ml. 30 μl of the samples were incubated at 40 °C for 1 week (Asn-deamidation) or 2 weeks (Asp-isomerization) in a heating block with a heated anti-condensation lid. The stressed and non-stressed samples were stored at -80 °C. Asn deamidation and Asp isomerization of the samples were analyzed by strong cation exchange (SCX) chromatography. In the SCX chromatogram, deamidation of Asn results in an increase in the peak area (bM) before the main peak, while in the SCX chromatogram, Asp isomerization results in an increase in the peak area (aM) after the main peak (Du et al. 2012). SCX chromatography was run on a MAbPac SCX-10 column (ThermoFisher Scientific, Basel, Switzerland) with buffer A at pH 4.0 and buffer B at pH 11.0. The flow rate was 0.5 ml / min and the pH gradient was 30 - 80% buffer B. 10 μg of the sample in 20 μl of buffer A was injected onto the column. Sample detection was performed by protein absorbance at 280 nm. The hCl antibody only showed an increase in bM of approximately 27.9 - 32.2% (see Table 6), which was rated as insignificant. However, IMAB362 showed a significant increase in bM of 40.9% (see Table 6), even though the antibody does not have an NG motif in the variable domain. Compared to the anti-CLDN18.2 monoclonal antibody of the present invention, IMAB362 has two NS motifs at positions HC CDR3 (amino acids 103 - 104) (SEQ ID NO:55) and LC CDR1 (amino acids 31 - 32) (SEQ ID NO:56). The NS motif is the second most likely motif for deamidation.

[0193] ​ Deamidation stress testing of mABs, strong cation exchange (SCX) chromatography

[0194]

[0195] The effect of Asn-deamidation stress testing on the binding affinity of hCl1a, hCl1i, and IMAB362 to CLDN18.2 was tested in an ELISA assay in which lipid particles bearing CLDN18.2 were used as the antigen source. CLDN18.2 lipid particles and empty lipid particles (antigen-free) were used to coat 96-well plates at a final concentration of 10 μg / ml in 100 mM sodium carbonate, pH 9.6. After washing with PBS / 0.05% Tween-20 (PBS-T) and blocking with PBS-T / 3% BSA at 37 °C for at least 1 hour, 1:3 serial dilutions of hCl antibodies at an initial concentration of 2 μg / ml were added and incubated at 37 °C for at least 1 hour. Color development was carried out by binding HRP-goat anti-human secondary antibody, with Sigma-Fast OPD as the peroxidase substrate, the reaction was terminated by adding 2 M H2SO4, and readings were taken at OD-490 on an ELISA plate reader, revealing the presence of bound antibody. After Asn-deamidation stress testing, the IMAB362 EC50 value was 1.8-fold higher (no stress treatment reference: EC50 = 51.5 ng / ml, stress treatment: EC50 = 95.09 ng / ml) (see ​ ). This may be related to the 40.9% increase in bM in SCX after Asn-deamidation stress testing (see Table 6). To confirm the SCX Asn-deamidation results, no significant differences in antigen binding were observed after Asn-deamidation stress testing of hCl1a and hCl1i (see Table 6). Thus, Asn-deamidation stress testing indicates that hCl antibodies are less prone to deamidation and potentially reduced target binding compared to IMAB362 and can be expected to be more stable during preparation, storage, and clinical application (in vivo), resulting in a more uniform and active antibody / product.

[0196] Although all hCl antibodies have potential DG Asp isomerization motifs in the 2nd CDR of VL and the CH2 and CH3 domains of HC (VL-CDR2 (position 62), CH2 (position 282), CH3 (position 403)), Asp isomerization stress testing did not show Asp isomerization (see Table 7), contrary to the prediction by Du et al. (Du et al. 2012). The aM values of samples without stress treatment (except IMAB362) were already significantly higher. This may be due to lysine clipping variants of the heavy chain. IMAB362 is the only antibody without high aM among samples without stress treatment. IMAB362 is the only anti-CLDN18.2 antibody tested without a C-terminal Lys, which means that for hCl antibodies, C-terminal Lys clipping is the most likely cause of the increase in aM in samples without stress treatment and stress treatment.

[0197] ​Asp Isomerization Stress Testing of mAb, Strong Cation Exchange (SCX) Chromatography

[0198]

[0199]

[0200] The present invention is also described by the following embodiments:

[0201] 1. An antibody or a fragment thereof that binds to CLDN18.2, wherein the antibody or the fragment thereof exhibits increased binding to tumor tissues expressing CLDN18.2 compared to healthy tissues expressing CLDN18.2.

[0202] 2. An antibody or a fragment thereof that binds to CLDN18.2, comprising HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:21, SEQ ID NO:22, and SEQ ID NO:23, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, respectively.

[0203] 3. The antibody or the fragment thereof according to embodiment 1 or 2, comprising:

[0204] a. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:15, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively;

[0205] b. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively;

[0206] c. HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3, respectively, and LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:17, SEQ ID NO:14, and SEQ ID NO:11, respectively;

[0207] d. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:16 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:11 respectively;

[0208] e. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:15 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively;

[0209] f. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively;

[0210] g. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20 and SEQ ID NO:3 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19 and SEQ ID NO:11 respectively;

[0211] h. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively; or

[0212] i. The HCDR1, HCDR2 and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO:17, SEQ ID NO:14 and SEQ ID NO:11 respectively.

[0213] 4. The antibody or fragment thereof of embodiment 1 or 2, comprising:

[0214] a. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively;

[0215] b. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:7, and SEQ ID NO:8 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11 respectively; or

[0216] c. The HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:2, and SEQ ID NO:3 respectively, and the LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:11 respectively.

[0217] 5. The antibody or fragment thereof of embodiment 1 or 2, which comprises:

[0218] a. The VH sequence of SEQ ID NO:27 and the VL sequence of SEQ ID NO:28;

[0219] b. The VH sequence of SEQ ID NO:29 and the VL sequence of SEQ ID NO:30; or

[0220] c. The VH sequence of SEQ ID NO:31 and the VL sequence of SEQ ID NO:32.

[0221] 6. The antibody or fragment thereof of any one of embodiments 1 - 3, which comprises:

[0222] a. A VH sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:33;

[0223] b. A VH sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:34;

[0224] c. A VH sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:35;

[0225] d. A VH sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:36; or

[0226] e. A VH sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:37;

[0227] and

[0228] f. A VL sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:38;

[0229] g. A VL sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:39;

[0230] h. A VL sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:40; or

[0231] i. A VL sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of SEQ ID NO:41.

[0232] 7. The antibody or fragment thereof according to embodiment 1 or 2, comprising:

[0233] a. The VH sequence of SEQ ID NO:33;

[0234] b. The VH sequence of SEQ ID NO:34;

[0235] c. The VH sequence of SEQ ID NO:35;

[0236] d. The VH sequence of SEQ ID NO:36; or

[0237] e. The VH sequence of SEQ ID NO:37;

[0238] and

[0239] f. The VL sequence of SEQ ID NO:38;

[0240] g. The VL sequence of SEQ ID NO:39;

[0241] h. The VL sequence of SEQ ID NO:40; or

[0242] i. The VL sequence of SEQ ID NO: 41.

[0243] 8. The antibody or fragment thereof according to embodiment 1 or 2, comprising:

[0244] a. The VH sequence of SEQ ID NO: 33 and the VL sequence of SEQ ID NO: 38;

[0245] b. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 38;

[0246] c. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 39;

[0247] d. The VH sequence of SEQ ID NO: 34 and the VL sequence of SEQ ID NO: 40;

[0248] e. The VH sequence of SEQ ID NO: 35 and the VL sequence of SEQ ID NO: 38;

[0249] f. The VH sequence of SEQ ID NO: 36 and the VL sequence of SEQ ID NO: 41;

[0250] g. The VH sequence of SEQ ID NO: 36 and the VL sequence of SEQ ID NO: 40;

[0251] h. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 41;

[0252] i. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 38; or

[0253] j. The VH sequence of SEQ ID NO: 37 and the VL sequence of SEQ ID NO: 39.

[0254] 9. The antibody according to any one of embodiments 1 - 3, comprising:

[0255] a. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the heavy chain sequence of SEQ ID NO: 46, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the light chain sequence of SEQ ID NO: 51;

[0256] b. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 47, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 51;

[0257] c. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 47, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 52;

[0258] d. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 47, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 53;

[0259] e. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 48, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 51;

[0260] f. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 47, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 54;

[0261] g. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the heavy chain sequence of SEQ ID NO: 49, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity with the amino acid sequence of the light chain sequence of SEQ ID NO: 53;

[0262] h. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the heavy chain sequence of SEQ ID NO:50, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the light chain sequence of SEQ ID NO:54;

[0263] i. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the heavy chain sequence of SEQ ID NO:50, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the light chain sequence of SEQ ID NO:51;

[0264] j. A heavy chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the heavy chain sequence of SEQ ID NO:50, and a light chain sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 98% sequence identity to the amino acid sequence of the light chain sequence of SEQ ID NO:52,

[0265] or an engineered Fc domain version thereof.

[0266] 10. The antibody of embodiment 1 or 2, comprising:

[0267] a. The heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51;

[0268] b. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:51;

[0269] c. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:52;

[0270] d. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:53;

[0271] e. The heavy chain sequence of SEQ ID NO:48 and the light chain sequence of SEQ ID NO:51;

[0272] f. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:54;

[0273] g. The heavy chain sequence of SEQ ID NO:49 and the light chain sequence of SEQ ID NO:53;

[0274] h. The heavy chain sequence of SEQ ID NO: 50 and the light chain sequence of SEQ ID NO: 54;

[0275] i. The heavy chain sequence of SEQ ID NO: 50 and the light chain sequence of SEQ ID NO: 51;

[0276] j. The heavy chain sequence of SEQ ID NO: 50 and the light chain sequence of SEQ ID NO: 52, or an engineered Fc domain version thereof.

[0277] 11. The antibody or fragment thereof according to any one of embodiments 1 to 10, wherein the antibody or fragment thereof is IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4, synthetic IgG, IgM, F(ab)2, Fv, scFv, IgGACH2, F(ab')2, scFvCH3, Fab, VL, VH, scFv4, scFv3, scFv2, dsFv, Fv, scFv-Fc, (scFv)2, non-consumable IgG, diabody, bivalent antibody or an engineered Fc version thereof.

[0278] 12. The antibody or fragment thereof according to any one of embodiments 1 to 11, wherein the antibody or fragment thereof is humanized.

[0279] 13. The antibody or fragment thereof according to any one of embodiments 1 to 12, wherein the antibody or fragment thereof does not bind to CLDN18.1.

[0280] 14. The antibody or fragment thereof according to any one of embodiments 1 to 13, wherein the antibody or fragment thereof is less sensitive to post-translational deamidation than IMAB362.

[0281] 15. The antibody or fragment thereof according to any one of embodiments 1 to 14, wherein the antibody or fragment thereof labels tumor cells expressing CLDN18.2 at least 2-fold, at least 5-fold, at least 10-fold or at least 20-fold more than healthy tissue cells expressing CLDN18.2 during flow cytometry measurement.

[0282] 16. The antibody or fragment thereof according to any one of embodiments 1 to 14, wherein the binding to tumor cells expressing CLDN18.2 is increased compared to healthy tissue cells expressing CLDN18.2 as measured by flow cytometry or by immunohistochemistry.

[0283] 17. An antibody or fragment thereof according to any one of embodiments 1 to 16, wherein the antibody or fragment thereof binds to CLDN18.2 expressed in HEK293T cells or PA-TU-8988-High with an EC50 value that is at least 1.1-fold, at least 1.2-fold, at least 1.5-fold, at least 2-fold, or at least 2.5-fold higher but not more than 3-fold higher than the EC50 value of IMAB362 binding to CLDN18.2 expressed in HEK293T cells or PA-TU-8988-High.

[0284] 18. The antibody or fragment thereof of embodiment 17, wherein the binding is measured by flow cytometry (FC) titration.

[0285] 19. An antibody or fragment thereof according to any one of embodiments 1 to 18, wherein the antibody or fragment thereof is isolated.

[0286] 20. A nucleic acid encoding an antibody or fragment thereof according to any one of embodiments 1 to 19.

[0287] 21. A vector comprising the nucleic acid of embodiment 20.

[0288] 22. A host cell comprising the nucleic acid of embodiment 20 or the vector of embodiment 21.

[0289] 23. An antibody or fragment thereof according to any one of embodiments 1 to 19, the nucleic acid of embodiment 20, the vector of embodiment 21, or the host cell of embodiment 22, for treating a subject:

[0290] a. suffering from a neoplastic disease,

[0291] b. at risk of developing a neoplastic disease, and / or

[0292] c. diagnosed with a neoplastic disease.

[0293] 24. The antibody or fragment thereof for embodiment 23, wherein the neoplastic disease is selected from pancreatic cancer, gastric cancer, esophageal cancer, ovarian cancer, and lung cancer.

[0294] 25. An antibody or fragment thereof that binds to CLDN18.2, wherein the antibody or fragment thereof

[0295] (i) binds to the same epitope as an antibody comprising the heavy chain sequence of SEQ ID NO: 46 and the light chain sequence of SEQ ID NO: 51;

[0296] (ii) competes for binding with an antibody comprising the heavy chain sequence of SEQ ID NO: 46 and the light chain sequence of SEQ ID NO: 51; and / or

[0297] (iii) Competitive inhibition of the binding of an antibody comprising the heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51 to CLDN18.2.

[0298] Sequence

[0299] SEQ ID NO:1 DYAMH

[0300] SEQ ID NO:2 WINTYTGKPTYADDFKG

[0301] SEQ ID NO:3 AVFYGYTMDA

[0302] SEQ ID NO:4 RASEDIYSNLA

[0303] SEQ ID NO:5 SVKRLQD

[0304] SEQ ID NO:6 LQGSNFPLT

[0305] SEQ ID NO:7 WINAYTGKPTYADDFKG

[0306] SEQ ID NO:8 AVYYGYTMDA

[0307] SEQ ID NO:9 RTSEDIYSNFA

[0308] SEQ ID NO:10 SVNRLQD

[0309] SEQ ID NO:11 LQGSKFPLT

[0310] SEQ ID NO:12 DYAMY

[0311] SEQ ID NO:13 RTSEDIYSNLA

[0312] SEQ ID NO:14 AIKRLQD

[0313] SEQ ID NO:15 WINTYTGKPTYAQKFQG

[0314] SEQ ID NO:16 WINTYTGKPTYSQKFQG

[0315] SEQ ID NO:17 RTSEDIYSNLA

[0316] SEQ ID NO:18 RTSEDIYSNFA

[0317] SEQ ID NO:19 SVNRLQD

[0318] SEQ ID NO:20 WINAYTGKPTYAQKFQG

[0319] SEQ ID NO:21 DYAMX

[0320] X at position 5 is H or Y

[0321] SEQ ID NO:22 WINXYTGKPTYXXXFXG

[0322] X at position 4 is T or A;

[0323] X at position 12 is A or S;

[0324] X at position 13 is D or Q;

[0325] X at position 14 is D or K;

[0326] X at position 16 is K or Q

[0327] SEQ ID NO:23 AVXYGYTMDA

[0328] X at position 3 is F or Y

[0329] SEQ ID NO:24 RXSEDIYSNXA

[0330] X at position 2 is A or T;

[0331] X at position 10 is L or F

[0332] SEQ ID NO:25 XXXRLQD

[0333] X at position 1 is S or A;

[0334] X at position 2 is V or I;

[0335] X at position 3 is K or N

[0336] SEQ ID NO:26 LQGSXFPLT

[0337] X at position 5 is K or N

[0338] SEQ ID NO:27 cCl1-1 HC variable region

[0339] QIQLVQSGPELKKPGESVKISCKASGYTFTDYAMHWVKQAPGKGLKWMGWINTYTGKPTYADDFKGRFVFSLEASASTANLQISNLKNEDTATYFCARAVFYGYTMDAWGQGTSVTVSS

[0340] SEQ ID NO:28 cCl1-1 LC variable region

[0341] DIQMTQSPASLSASLGETISIACRASEDIYSNLAWYQQKSGKSPQLLIFSVKRLQDGVPSRFSGSGSGTQYSLKISGMQPEDEGDYFCLQGSNFPLTFGSGTKLEIK

[0342] SEQ ID NO:29 cCl1-2 HC variable region

[0343] QIQLVQSGPELKKPGESVKISCKTSGYTFTDYAMHWVKQGPGKGMKWMGWINAYTGKPTYADDFKGRFVLSLEASASTANLQISNLKNEDTATYFCARAVYYGYTMDAWGQGTSVIVSS

[0344] SEQ ID NO:30 cCl1-2 LC variable region

[0345] DIQMTQSPASLSASLGETISIECRTSEDIYSNFAWFQQKSGKSPQLLIYSVNRLQDGVPSRFSGSGSGTQYSLKISGMQPEDEGDYFCLQGSKFPLTFGSGTKLEIK

[0346] SEQ ID NO:31 cCl1-3 HC variable region

[0347] QIQLVQSGPELKKPGESVKISCKASGYTFTDYAMYWVKQVPGKGLRWMGWINTYTGKPTYADDFKGRFVFSLEASASTANLQISNLKNEDTATYFCARAVFYGYTMDAWGQGTSVTVSS

[0348] SEQ ID NO:32 cCl1-3 LC variable region

[0349] DIQMTQSPASLSASLGETISIACRTSEDIYSNLAWYQQKSGKSPQLLIFAIKRLQDGVPSRFSGSGSGTQYSLKISGMQPEDEGDYFCLQGSKFPLTFGSGTKLEIK

[0350] SEQ ID NO:33 hCL1a HC variable region

[0351] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINTYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSS

[0352] SEQ ID NO:34 hCL1b, c and d HC variable region

[0353] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINTYTGKPTYSQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSS

[0354] SEQ ID NO:35 hCL1e HC variable region

[0355] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMYWVRQAPGQRLEWMGWINTYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSS

[0356] SEQ ID NO:36 hCL1f and g HC variable region

[0357] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINAYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSS

[0358] SEQ ID NO:37 hCL1h, i and j HC variable region

[0359] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMYWVRQAPGQRLEWMGWINAYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVYYGYTMDAWGQGTLVTVSS

[0360] SEQ ID NO:38 hCL1a, b, e and i LC variable regions

[0361] DIQMTQSPSSLSASVGDRVTITCRASEDIYSNLAWYQQKPGKAPKLLIFSVKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSNFPLTFGQGTKVEIK

[0362] SEQ ID NO:39 hCL1c and j LC variable regions

[0363] DIQMTQSPSSLSASVGDRVTITCRTSEDIYSNLAWYQQKPGKAPKLLIFAIKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSKFPLTFGQGTKVEIK

[0364] SEQ ID NO:40 hCL1d and g LC variable regions

[0365] DIQMTQSPSSLSASVGDRVTITCRTSEDIYSNFAWYQQKPGKAPKLLIYSVNRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSKFPLTFGQGTKVEIK

[0366] SEQ ID NO:41 hCL1f and h LC variable regions

[0367] DIQMTQSPSSLSASVGDRVTITCRASEDIYSNLAWYQQKPGKAPKLLIYSVKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSNFPLTFGQGTKVEIK

[0368] SEQ ID NO:42 hCL3a, b and c HC variable regions

[0369] QVQLQESGPGLVKPSETLSLTCAVSGYSVSSNYRWHWIRQPPGKGLEWIGYINIAGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARNPSITRAMDAWGQGTLVTVSS

[0370] SEQ ID NO:43 Variable region of hCL3a LC

[0371] DIQMTQSPSSLSASVGDRVTITCKSSQNIFKNLEWYQQKPGKAPKLLIYYTNNLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCYQYNSGPFTFGQGTKVEIK

[0372] SEQ ID NO:44 Variable region of hCL3b LC

[0373] DIQMTQSPSSLSASVGDRVTITCRSSQNIFKNLEWYQQKPGKAPKLLIYYTNNLQTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCYQYNSGPFTFGQGTKVEIK

[0374] SEQ ID NO:45 Variable region of hCL3c LC

[0375] DIQMTQSPSSLSASVGDRVTITCRSSQNIFKNLEWYQQKPGKAPKLLIYYTNNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCYQYNSGPFTFGQGTKVEIK

[0376] SEQ ID NO:46 Full-length hCL1a HC

[0377] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINTYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0378] SEQ ID NO:47 hCL1b, c and d HC full length

[0379] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINTYTGKPTYSQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0380] SEQ ID NO:48 Full-length hCL1e HC

[0381] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMYWVRQAPGQRLEWMGWINTYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0382] SEQ ID NO:49 Full-length hCL1f and g HC

[0383] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMHWVRQAPGQRLEWMGWINAYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVFYGYTMDAWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0384] SEQ ID NO:50 hCL1 h, i and j HC full length

[0385] QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYAMYWVRQAPGQRLEWMGWINAYTGKPTYAQKFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARAVYYGYTMDAWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0386] SEQ ID NO:51 Full-length hCL1a, b, e and i LC

[0387] DIQMTQSPSSLSASVGDRVTITCRASEDIYSNLAWYQQKPGKAPKLLIFSVKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSNFPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0388] SEQ ID NO:52 Full-length hCL1c and j LC

[0389] DIQMTQSPSSLSASVGDRVTITCRTSEDIYSNLAWYQQKPGKAPKLLIFAIKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSKFPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0390] SEQ ID NO:53 Full-length hCL1d and g LC

[0391] DIQMTQSPSSLSASVGDRVTITCRTSEDIYSNFAWYQQKPGKAPKLLIYSVNRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSKFPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0392] SEQ ID NO:54 Full-length hCL1f and h LC

[0393] DIQMTQSPSSLSASVGDRVTITCRASEDIYSNLAWYQQKPGKAPKLLIYSVKRLQDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQGSNFPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0394] SEQ ID NO:55 IMAB362 HC full length

[0395] QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0396] SEQ ID NO:56 IMAB362 LC full length

[0397] DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0398] SEQ ID NO:57 DQWSTQDLYN

[0399] SEQ ID NO:58 NNPVTAVFNYQ

[0400] SEQ ID NO:59 STQDLYNNPVTAVF

[0401] SEQ ID NO:60 TNFWMSTANMYTG

[0402] SEQ ID NO:61 ALMIVGIVLGAIGLLV

[0403] SEQ ID NO:62 RIGSMEDSAKANMTLTSGIMFIVS

[0404] SEQ ID NO:63

[0405] METDTLLLWVLLLWVPGSTGDAAQPARRARRTKLGTELGSTPVWWNSADGRMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRAAIQHSGGRSRRARTKTHLRRGSE

[0406] SEQ ID NO:64 MDQWSTQDLYNNPVT

[0407] SEQ ID NO:65 LYNNPVTAVFNYQGL

[0408] SEQ ID NO:66 VFNYQGLWRSCVRES

[0409] SEQ ID NO:67 QGLWRSCVRESSGFT

[0410] SEQ ID NO:68 RSCVRESSGFTECRG

[0411] SEQ ID NO:69 TEDEVQSYPSKHDYV

[0412] SEQ ID NO:70 EVQSYPSKHDYV

[0413] SEQ ID NO:71 gactacgcgatgcac

[0414] SEQ ID NO:72 tggatcaacacgtacacggggaagccgacatacgcggacgacttcaagggg

[0415] SEQ ID NO:73 gccgtcttctacggatatacgatggacgcg

[0416] SEQ ID NO:74

[0417] cagatccagctcgtccagagcgggccggagctgaagaagccgggggagagcgtgaagatctcgtgcaaggcgagcggatatacgttcacggactacgcgatgcactgggtcaagcaagcgccggggaaagggctgaagtggatggggtggatcaacacgtacacggggaagccgacatacgcggacgacttcaaggggcgattcgtgttctcgctggaggcgagcgcgagcacggcgaacctgcaaatctcgaacctgaagaacgaggacacggcgacgtacttctgcgcgcgggccgtcttctacggatatacgatggacgcgtgggggcagggtaccagcgtgacggtctcgagc

[0418] SEQ ID NO:75 cgggcgagcgaggacatctactcgaacctggcg

[0419] SEQ ID NO:76 tccgtcaagcggctgcaagac

[0420] SEQ ID NO:77 ctgcaagggagcaacttcccgctgacg

[0421] SEQ ID NO:78

[0422] gacatccagatgacgcagagcccggcgtcgctgagcgcgagcctgggggagacgatctcgatcgcgtgccgggcgagcgaggacatctactcgaacctggcgtggtatcaacagaagagcgggaagagcccgcagctgctgatcttctccgtcaagcggctgcaagacggcgtcccgagccgattctcggggagcgggagcgggacgcagtactcgctgaagatctcggggatgcagccggaggacgagggggactacttctgcctgcaagggagcaacttcccgctgacgttcgggtcgggtaccaaactcgagatcaaa gatccagatgacgcagagcccggcgtcgctgagcgcgagcctgggggagacgatctcgatcgcgtgccgggcgagcgaggacatctactcgaacctggcgtggtatcaacagaagagcgggaagagcccgcagctgctgatcttctccgtcaagcggctgcaagacggcgtcccgagccgattctcggggagcgggagcgggacgcagtactcgctgaagatctcggggatgcagccggaggacgagggggactacttctgcctgcaagggagcaacttcccgctgacgttcgggtcgggtaccaaactcgagatcaaa

[0423] SEQ ID NO:79 tggatcaacgcgtacacggggaagccgacctacgcggacgacttcaagggg tggatcaacgcgtacacggggaagccgacctacgcggacgacttcaagggg

[0424] SEQ ID NO:80 gccgtctactacggatatacgatggac gccgtctactacggatatacgatggac

[0425] SEQ ID NO:81

[0426] cagatccagctcgtccagagcgggccggagctgaagaagccgggggagagcgtgaagatctcgtgcaagacgagcggatatacgttcacggactacgcgatgcactgggtcaagcaggggccagggaaagggatgaagtggatggggtggatcaacgcgtacacggggaagccgacctacgcggacgacttcaaggggcgattcgtgctgagcctggaggcgagcgcctcgacggcgaacctgcaaatctcgaacctgaagaacgaggacacggcgacgtacttctgcgcgcgggccgtctactacggatatacgatggacgcgtgggggcagggtaccagcgtgatcgtctcgagc cagatccagctcgtccagagcgggccggagctgaagaagccgggggagagcgtgaagatctcgtgcaagacgagcggatatacgttcacggactacgcgatgcactgggtcaagcaggggccagggaaagggatgaagtggatggggtggatcaacgcgtacacggggaagccgacctacgcggacgacttcaaggggcgattcgtgctgagcctggaggcgagcgcctcgacggcgaacctgcaaatctcgaacctgaagaacgaggacacggcgacgtacttctgcgcgcgggccgtctactacggatatacgatggacgcgtgggggcagggtaccagcgtgatcgtctcgagc

[0427] SEQ ID NO:82 cggacgagcgaggacatctactcgaacttcgcg

[0428] SEQ ID NO:83 tcagtcaaccggctgcaagac

[0429] SEQ ID NO:84 ctgcaagggagcaagttcccgctgacg

[0430] SEQ ID NO:85

[0431] gacatccagatgacgcagagcccggcgagcctgagcgcgagcctgggggagacgatctcgatcgagtgccggacgagcgaggacatctactcgaacttcgcgtggttccagcagaagagcgggaagagcccgcagctgctgatctactcagtcaaccggctgcaagacggcgtcccgagccgattctcggggagcgggagcgggacgcagtactcgctgaagatctcggggatgcagccggaggacgagggggactacttctgcctgcaagggagcaagttcccgctgacgttcgggagcggtaccaaactcgagatcaaa

[0432] SEQ ID NO:86 gactacgcgatgtac

[0433] SEQ ID NO:87 tggatcaacacgtacacggggaagccgacctacgcggacgacttcaagggg

[0434] SEQ ID NO:88

[0435] cagatccagctcgtccagagcgggccggagctgaagaagccgggggagagcgtgaagatctcgtgcaaggcgagcggatatacgttcacggactacgcgatgtactgggtcaagcaagtgccggggaaagggctgcgatggatggggtggatcaacacgtacacggggaagccgacctacgcggacgacttcaaggggcgattcgtgttctcgctggaggcgagcgcgagcacggcgaacctgcaaatctcgaacctgaagaacgaggacacggcgacgtacttctgcgcgcgggccgtcttctacggatatacgatggacgcgtgggggcagggtaccagcgtgacggtctcgagc

[0436] SEQ ID NO:89 cggacgagcgaggacatctactcgaacctggcg

[0437] SEQ ID NO:90 gcgatcaagcggctgcaagac

[0438] SEQ ID NO:91

[0439] gacatccagatgacgcagagcccggcgagcctgagcgcgagcctgggggagacgatctcgatcgcgtgccggacgagcgaggacatctactcgaacctggcgtggtatcaacagaagagcgggaagagcccgcagctgctgatcttcgcgatcaagcggctgcaagacggcgtcccgagccgattctcggggagcgggagcgggacgcagtactcgctgaagatctcggggatgcagccggaggacgagggggactacttctgcctgcaagggagcaagttcccgctgacgttcgggtcgggtaccaaactcgagatcaaa

[0440] SEQ ID NO:92 tggatcaatacatacacggggaagccgacttatgcgcaaaaattccaagga

[0441] SEQ ID NO:93 gcggtcttctacggatatacgatggatgcc

[0442] SEQ ID NO:94

[0443] caggtccaactagtccaaagcggggcggaagtcaagaagcccggagcatccgtcaaagtcagctgcaaggcgagcggatatacatttacggactacgcgatgcactgggtcaggcaagcccctgggcaaaggctcgaatggatgggatggatcaatacatacacggggaagccgacttatgcgcaaaaattccaaggaagagtcacaattacgcgggatacatccgcatctaccgcctacatggagctaagctcgctgcggagcgaggatacggcggtctactattgcgcccgagcggtcttctacggatatacgatggatgcctgggggcagggtaccctggtcacggtctcgagc

[0444] SEQ ID NO:95 agggcctccgaagacatctactccaacctggca

[0445] SEQ ID NO:96 agcgtcaaaagactacaagat

[0446] SEQ ID NO:97 ttgcaaggaagcaatttccccttgact

[0447] SEQ ID NO:98

[0448] gacattcaaatgacgcaaagcccatcatcgctgagcgcatcggtcggggatagagtcaccataacatgcagggcctccgaagacatctactccaacctggcatggtatcaacaaaaaccggggaaggctccgaagctgctgatatttagcgtcaaaagactacaagatggagtaccgagccgattttcgggaagcgggagcgggacggatttcacgctgaccatatcaagtttgcaaccggaggattttgcgacatactattgcttgcaaggaagcaatttccccttgactttcgggcaaggtaccaaggtcgagatcaaa

[0449] SEQ ID NO:99 gattatgcaatgcac

[0450] SEQ ID NO:100 tggattaacacctacacgggcaagcccacatactcccaaaaattccaagga

[0451] SEQ ID NO:101 gctgtattctatggatatacaatggatgcc

[0452] SEQ ID NO:102

[0453] caggtccaattagtccaaagcggggcggaagtcaagaagccgggggcgagcgtcaaagtctcatgcaaagcgagcggatacacatttacggattatgcaatgcactgggtcaggcaagcacccggacaaaggctggaatggatgggatggattaacacctacacgggcaagcccacatactcccaaaaattccaaggaagggtcacgataacgagagacacgagcgcgagcaccggaatggatgggatggattaacacctacacgggcaagcccacatactcccaaaaattccaaggaagggtcacgataacgagagacacgagcgcgagcaccgtaccctggtcaccgtctcgagc

[0454] SEQ ID NO:103 cgaacgagcgaggacatatactcaaaccttgca

[0455] SEQ ID NO:104 gcgataaagaggctgcaagac

[0456] SEQ ID NO:105 ttgcaaggctccaaatttcccctgaca

[0457] SEQ ID NO:106

[0458] gacatccaaatgactcaaagcccatcatcgctatcggcatcggtcggggatagagtcacgataacatgccgaacgagcgaggacatatactcaaaccttgcatggtatcaacaaaagccggggaaggccccgaagctactgatattcgcgataaagaggctgcaagacggagttccatcacgattttcgggatctggctcggggaccgattttacgctgactatatcatcgctgcaaccggaagattttgcaacatactactgcttgcaaggctccaaatttcccctgacattcggacaaggtaccaaggtcgagatcaaa

[0459] SEQ ID NO:107 cggacgagcgaggatatttattcgaactttgca

[0460] SEQ ID NO:108 cagtcaatcggctacaagat

[0461] SEQ ID NO:109

[0462] gacatccaaatgacgcaatcaccgagctcgctgagcgcatctgtcggggaccgtgtcacaatcacatgccggacgagcgaggatatttattcgaactttgcatggtatcaacaaaaaccgggcaaggctccgaaacttttgatttattcagtcaatcggctacaagatggcgtcccgagccgatttagcgggagcggatcgggaaccgactttacgctgacgatatcatcgctacaaccggaggacttcgcgacttattactgcctacaagggagcaaattcccgctgacattcggacaaggtaccaaggtcgagatcaaa

[0463] SEQ ID NO:110 gattacgcaatgtac

[0464] SEQ ID NO:111 tggataaatacctatacgggaaagccaacatacgcccaaaaattccaaggc

[0465] SEQ ID NO:112 gccgtcttttatggatatacgatggacgca

[0466] SEQ ID NO:113

[0467] caggtccaactggtccaatcgggggctgaagtcaaaaagccgggggcgagcgtcaaagtcagctgcaaagcatcgggatacacatttacggattacgcaatgtactgggtcaggcaagcacccggccaacgactggaatggatgggctggataaatacctatacgggaaagccaacatacgcccaaaaattccaaggccgcgtcacaataacgcgggacacgagcgcatcgacggcttatatggaactatcatcgctgcgatcggaagacacggcggtctattattgcgcacgcgccgtcttttatggatatacgatggacgcatgggggcagggtaccctggtcacggtctcgagc

[0468] SEQ ID NO:114 gactacgcaatgcac

[0469] SEQ ID NO:115 tggattaatgcctacacggggaagccgacctacgcacaaaaattccaagga

[0470] SEQ ID NO:116 gccgtcttctatggatatacgatggatgct

[0471] SEQ ID NO:117

[0472] caggtccaattggtccaaagcggggcggaggtcaagaagccgggggcgagcgtcaaagtctcatgcaaggcaagcggatatacatttacggactacgcaatgcactgggtccggcaagcccctgggcaacggctggaatggatgggatggattaatgcctacacggggaagccgacctacgcacaaaaattccaaggacgagtcacgattacgcgggatactagcgcgagcaccgcatatatggagctaagctcgctgcgatctgaggataccgctgtatactactgcgcgagagccgtcttctatggatatacgatggatgcttgggggcagggtaccctggtcacggtctcgagc

[0473] SEQ ID NO:118 cgagcttcggaggacatctatagcaacttggct

[0474] SEQ ID NO:119 agcgtcaaaaggctccaagac

[0475] SEQ ID NO:120 ctacaaggctctaacttcccattgaca

[0476] SEQ ID NO:121

[0477] gatatccaaatgacgcaatcaccatctagcctatcggcctctgtgggggaccgagtcaccatcacatgccgagcttcggaggacatctatagcaacttggcttggtatcaacaaaagccggggaaagcaccaaagctgctgatatatagcgtcaaaaggctccaagacggagtcccaagccgattctcgggctccggctccgggacggattttacgctgacaatttcgagcctgcaaccggaggactttgcaacctactattgcctacaaggctctaacttcccattgacatttgggcaaggtaccaaggtcgagatcaaa

[0478] SEQ ID NO:122 gactacgctatgtat

[0479] SEQ ID NO:123 tggattaatgcctacaccgggaagccgacttatgcgcaaaaatttcaagga

[0480] SEQ ID NO:124 gcggtctactatggatatacgatggacgca

[0481] SEQ ID NO:125

[0482] caggtccaactggttcaatctggagcggaagtcaagaagcccggagcatccgtcaaagtctcgtgcaaggcatctggatacacattcaccgactacgctatgtattgggtccggcaagcccccggacaacggctggaatggatgggatggattaatgcctacaccgggaagccgacttatgcgcaaaaatttcaaggaagggtcacgattacgcgggacacgagcgcctcaaccgcatacatggagctatcgagcctgcgaagcgaggacaccgcggtctactactgcgcgcgggcggtctactatggatatacgatggacgcatgggggcagggtaccctggtcacggtctcgagc

[0483] SEQ ID NO:126 WINXYTGKPTYXQKFQG

[0484] X at the 4th position is T or A;

[0485] X at the 12th position is A or S

[0486] [HC CDR2 is only for hCl1x, not for chimeric clones cCl1-1,2,3]

[0487] SEQ ID NO:127

[0488] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC [Constant light chain - CL domain]

[0489] SEQ ID NO:128

[0490] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK [Constant heavy chain - CH1 + Fc domain]

[0491] SEQ ID NO:129

[0492] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK [L234A / L235A mutation in the constant heavy chain - CH1+Fc domain]

[0493] SEQ ID NO:130

[0494] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK [L236A / L236A / P329G mutation in the constant heavy chain - CH1+Fc domain]

[0495] SEQ ID NO:131 RLPQTGG [Sortase tag]

[0496] SEQ ID NO:132 GGGGS-LPQTGG [Sortase tag]

[0497] SEQ ID NO:133 CLDN18.2

[0498] MAVTACQGLGFVVSLIGIAGIIAATCMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRALMIVGIVLGAIGLLVSIFALKCIRIGSMEDSAKANMTLTSGIMFIVSGLCAIAGVSVFANMLVTNFWMSTANMYTGMGGMVQTVQTRYTFGAALFVGWVAGGLTLIGGVMMCIACRGLAPEETNYKAVSYHASGHSVAYKPGGFKASTGFGSNTKNKKIYDGGARTEDEVQSYPSKHDYV

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[0547] Zalevsky, J., A. K. Chamberlain, H. M. Horton, S. Karki, I. W. Leung, T. J. Sproule, G. A. Lazar, D. C. Roopenian, and J. R. Desjarlais. 2010. 'Enhanced antibody half - life improves in vivo activity', Nat Biotechnol, 28:157 - 9.

[0548] CN109762067

[0549] WO2000 / 015659

[0550] WO2004 / 047863

[0551] WO2005 / 113587

[0552] WO2007 / 059997

[0553] WO2008 / 145338

[0554] WO2013 / 167259

[0555] WO2013 / 174509

[0556] WO2014 / 075788

[0557] WO2014 / 127906

[0558] WO2016 / 166122

[0559] WO2016 / 165762

[0560] WO2018 / 006882

[0561] WO2019 / 173420

[0562] WO2019 / 175617

[0563] WO2019 / 219089 Sequence Listing <110> Codiak BioSciences, Inc. <120> Tumor-Specific Claudin 18.2 Antibody <130> S12411WO / SOTCLD-1904 <150> 19 219 359.7 <151> 2019-12-23 <150> 20 152 510.2 <151> 2020-01-17 <160> 133 <170> PatentIn version 3.5 <210> 1 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, cCl1-2, hCl1a, hCl1b, hCl1c, hCl1d, hCl1f, hCl1g HCDR1 <400> 1 Asp Tyr Ala Met His 1 5 <210> 2 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, cCl1-3 HCDR2 <400> 2 Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 3 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, cCl1-3 , hCl1a, hCl1b, hCl1c, hCl1d, hCl1e, hCl1f,hCl1g HCDR3 <400> 3 Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala 1 5 10 <210> 4 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, hCl1a, hCl1b, hCl1e, hCl1f, hCl1h, hCl1i LCDR1 <400> 4 Arg Ala Ser Glu Asp Ile Tyr Ser Asn Leu Ala 1 5 10 <210> 5 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, hCl1a, hCl1b, hCl1e, hCl1f, hCl1h, hCl1i LCDR2 <400> 5 Ser Val Lys Arg Leu Gln Asp 1 5 <210> 6 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1, hCl1a, hCl1b, hCl1e, hCl1f, hCl1h, hCl1i LCDR3 <400> 6 Leu Gln Gly Ser Asn Phe Pro Leu Thr 1 5 <210> 7 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> cCl1-2 HCDR2 <400> 7 Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 8 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> cCl1-2, hCl1h, hCl1i, hCl1j HCDR3 <400> 8 Ala Val Tyr Tyr Gly Tyr Thr Met Asp Ala 1 5 10 <210> 9 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> cCl1-2 LCDR1 <400> 9 Arg Thr Ser Glu Asp Ile Tyr Ser Asn Phe Ala 1 5 10 <210> 10 <211> 7 <212> PRT <213> Artificial sequence <220> <223> cCl1-2 LCDR2 <400> 10 Ser Val Asn Arg Leu Gln Asp 1 5 <210> 11 <211> 9 <212> PRT <213> Artificial sequence <220> <223> cCl1-2, cCl1-3, hCl1c, hCl1d, hCl1g, hCl1j LCDR3 <400> 11 Leu Gln Gly Ser Lys Phe Pro Leu Thr 1 5 <210> 12 <211> 5 <212> PRT <213> Artificial sequence <220> <223> cCl1-3, hCl1e, hCl1h, hCl1i, hCl1j HCDR1 <400> 12 Asp Tyr Ala Met Tyr 1 5 <210> 13 <211> 11 <212> PRT <213> Artificial sequence <220> <223> cCl1-3 LCDR1 <400> 13 Arg Thr Ser Glu Asp Ile Tyr Ser Asn Leu Ala 1 5 10 <210> 14 <211> 7 <212> PRT <213> Artificial sequence <220> <223> cCl1-3, hCl1c, hCl1j LCDR2 <400> 14 Ala Ile Lys Arg Leu Gln Asp 1 5 <210> 15 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> hCl1a, hCl1e HCDR2 <400> 15 Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 16 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> hCl1b, hCl1c, hCl1d HCDR2 <400> 16 Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ser Gln Lys Phe Gln 1 5 10 15 Gly <210> 17 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> hCl1c, hCl1j LCDR1 <400> 17 Arg Thr Ser Glu Asp Ile Tyr Ser Asn Leu Ala 1 5 10 <210> 18 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> hCl1d, hCl1g LCDR1 <400> 18 Arg Thr Ser Glu Asp Ile Tyr Ser Asn Phe Ala 1 5 10 <210> 19 <211> 7 <212> PRT <213> Artificial sequence <220> <223> hCl1d, hCl1g LCDR2 <400> 19 Ser Val Asn Arg Leu Gln Asp 1 5 <210> 20 <211> 17 <212> PRT <213> Artificial sequence <220> <223> hCl1f, hCl1g, hCl1h, hCl1i, hCl1j HCDR2 <400> 20 Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 21 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Consensus sequence of HCDR1 <220> <221> Miscellaneous feature <222> (5)..(5) <223> H or Y <400> 21 Asp Tyr Ala Met Xaa 1 5 <210> 22 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Consensus HCDR2 sequence <220> <221> Miscellaneous features <222> (4)..(4) <223> T or A <220> <221> Miscellaneous features <222> (12)..(12) <223> A or S <220> <221> Miscellaneous features <222> (13)..(13) <223> D or Q <220> <221> Miscellaneous features <222> (14)..(14) <223> D or K <220> <221> Miscellaneous features <222> (16)..(16) <223> K or Q <400> 22 Trp Ile Asn Xaa Tyr Thr Gly Lys Pro Thr Tyr Xaa Xaa Xaa Phe Xaa 1 5 10 15 Gly <210> 23 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Consensus HCDR3 sequence <220> <221> Miscellaneous features <222> (3)..(3) <223> F or Y <400> 23 Ala Val Xaa Tyr Gly Tyr Thr Met Asp Ala 1 5 10 <210> 24 <211> 11 <212> PRT <213> Artificial sequence <220> <223> LCDR1 consensus sequence <220> <221> Miscellaneous features <222> (2)..(2) <223> A or T <220> <221> Miscellaneous features <222> (10)..(10) <223> L or F <400> 24 Arg Xaa Ser Glu Asp Ile Tyr Ser Asn Xaa Ala 1 5 10 <210> 25 <211> 7 <212> PRT <213> Artificial sequence <220> <223> LCDR2 consensus sequence <220> <221> Miscellaneous features <222> (1)..(1) <223> S or A <220> <221> Miscellaneous features <222> (2)..(2) <223> V or I <220> <221> Miscellaneous features <222> (3)..(3) <223> K or N <400> 25 Xaa Xaa Xaa Arg Leu Gln Asp 1 5 <210> 26 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Consensus sequence of LCDR3 <220> <221> Miscellaneous features <222> (5)..(5) <223> K or N <400> 26 Leu Gln Gly Ser Xaa Phe Pro Leu Thr 1 5 <210> 27 <211> 119 <212> PRT <213> Artificial sequence <220> <223> cCl1-1 HC variable region <400> 27 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Glu Ala Ser Ala Ser Thr Ala Asn 65 70 75 80 Leu Gln Ile Ser Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 28 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> cCl1-1 LC Variable Region <400> 28 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Glu Thr Ile Ser Ile Ala Cys Arg Ala Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Ser Gly Lys Ser Pro Gln Leu Leu Ile 35 40 45 Phe Ser Val Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Ser Gly Met Gln Pro 65 70 75 80 Glu Asp Glu Gly Asp Tyr Phe Cys Leu Gln Gly Ser Asn Phe Pro Leu 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 29 <211> 119 <212> PRT <213> Artificial sequence <220> <223> cCl1-2 HC variable region <400> 29 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Lys Gln Gly Pro Gly Lys Gly Met Lys Trp Met 35 40 45 Gly Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Leu Ser Leu Glu Ala Ser Ala Ser Thr Ala Asn 65 70 75 80 Leu Gln Ile Ser Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Ala Val Tyr Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Ser Val Ile Val Ser Ser 115 <210> 30 <211> 107 <212> PRT <213> Artificial sequence <220> <223> cCl1-2 LC variable region <400> 30 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Glu Thr Ile Ser Ile Glu Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Phe Ala Trp Phe Gln Gln Lys Ser Gly Lys Ser Pro Gln Leu Leu Ile 35 40 45 Tyr Ser Val Asn Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Ser Gly Met Gln Pro 65 70 75 80 Glu Asp Glu Gly Asp Tyr Phe Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 31 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> cCl1-3 HC variable region <400> 31 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met Tyr Trp Val Lys Gln Val Pro Gly Lys Gly Leu Arg Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Val Phe Ser Leu Glu Ala Ser Ala Ser Thr Ala Asn 65 70 75 80 Leu Gln Ile Ser Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 32 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> cCl1-3 LC variable region <400> 32 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Glu Thr Ile Ser Ile Ala Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Ser Gly Lys Ser Pro Gln Leu Leu Ile 35 40 45 Phe Ala Ile Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Ser Gly Met Gln Pro 65 70 75 80 Glu Asp Glu Gly Asp Tyr Phe Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 33 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl1a HC Variable Region <400> 33 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 34 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl1b, hCl1c and hCl1d HC Variable Regions <400> 34 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ser Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 35 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl1e HC Variable Region <400> 35 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 36 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl1f and hCl1g HC Variable Regions <400> 36 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 37 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl1h, hCl1i and hCl1j HC Variable Regions <400> 37 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Tyr Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 38 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Variable regions of hCl1a, hCl1b, hCl1e and hCl1i LC <400> 38 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ser Val Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Asn Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 39 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Variable regions of hCl1c and hCl1j LC <400> 39 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ala Ile Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 40 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> hCl1d and hCl1g LC Variable Regions <400> 40 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Phe Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Val Asn Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 41 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> hCl1f and hCl1h LC variable regions <400> 41 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Val Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Asn Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 42 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> hCl3a, hCl1b and hCl1c HC Variable Regions <400> 42 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Tyr Ser Val Ser Ser Asn 20 25 30 Tyr Arg Trp His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Asn Ile Ala Gly Ser Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Pro Ser Ile Thr Arg Ala Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 43 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> hCl3a LC Variable Region <400> 43 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Asn Ile Phe Lys Asn 20 25 30 Leu Glu Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Asn Asn Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Tyr Gln Tyr Asn Ser Gly Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 44 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> hCl3b LC Variable Region <400> 44 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Asn Ile Phe Lys Asn 20 25 30 Leu Glu Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Asn Asn Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Tyr Gln Tyr Asn Ser Gly Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 45 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> hCl3c LC variable region <400> 45 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Asn Ile Phe Lys Asn 20 25 30 Leu Glu Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Asn Asn Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Tyr Gln Tyr Asn Ser Gly Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 46 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> hCl1a HC full length <400> 46 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 47 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> Full-length hCl1b, hCl1c and hCl1d HC <400> 47 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ser Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 48 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> hCl1e HC Full Length <400> 48 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 49 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> hCl1f and hCl1g HC full length <400> 49 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Phe Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 50 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> Full length of hCl1h, hCl1i and hCl1j HC <400> 50 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met 35 40 45 Gly Trp Ile Asn Ala Tyr Thr Gly Lys Pro Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Val Tyr Tyr Gly Tyr Thr Met Asp Ala Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 51 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Full-length hCl1a, hCl1b, hCl1e and hCl1i LC <400> 51 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ser Val Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Asn Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 52 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> Full length hCl1c and hCl1j LC <400> 52 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Phe Ala Ile Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 53 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> hCl1d and hCl1g LC Full Length <400> 53 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Phe Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Val Asn Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Lys Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 54 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> hCl1f and hCl1h LC full length <400> 54 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Val Lys Arg Leu Gln Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Gly Ser Asn Phe Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 55 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> IMAB362 HC Full Length <400> 55 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 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 56 <211> 220 <212> PRT <213> Artificial Sequence <220> <223> IMAB362 LC Full Length <400> 56 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 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 57 <211> 10 <212> PRT <213> Artificial sequence <220> <223> The N-terminal extracellular region of CLDN18.2, not related to glycosylation <400> 57 Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn 1 5 10 <210> 58 <211> 11 <212> PRT <213> Artificial sequence <220> <223> The N-terminal extracellular region of CLDN18.2, largely unglycosylated <400> 58 Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln 1 5 10 <210> 59 <211> 14 <212> PRT <213> Artificial sequence <220> <223> The N-terminal extracellular domain of CLDN18.2, unglycosylated <400> 59 Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe 1 5 10 <210> 60 <211> 13 <212> PRT <213> Artificial sequence <220> <223> Pan - CLDN18 peptide in the C - terminal extracellular domain shared by CLDN18.1 and CLDN18.2 subtypes <400> 60 Thr Asn Phe Trp Met Ser Thr Ala Asn Met Tyr Thr Gly 1 5 10 <210> 61 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Specific epitope of CLDN18.2 as disclosed in WO2005 / 113587 <400> 61 Ala Leu Met Ile Val Gly Ile Val Leu Gly Ala Ile Gly Leu Leu Val 1 5 10 15 <210> 62 <211> 24 <212> PRT <213> Artificial sequence <220> <223> Specific epitope of CLDN18.2 as disclosed in WO2005 / 113587 <400> 62 Arg Ile Gly Ser Met Glu Asp Ser Ala Lys Ala Asn Met Thr Leu Thr 1 5 10 15 Ser Gly Ile Met Phe Ile Val Ser 20 <210> 63 <211> 129 <212> PRT <213> Artificial sequence <220> <223> The first extracellular domain of CLDN18.2, with N-terminal and C-terminal extensions <400> 63 Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 1 5 10 15 Gly Ser Thr Gly Asp Ala Ala Gln Pro Ala Arg Arg Ala Arg Arg Thr 20 25 30 Lys Leu Gly Thr Glu Leu Gly Ser Thr Pro Val Trp Trp Asn Ser Ala 35 40 45 Asp Gly Arg Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro 50 55 60 Val Thr Ala Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg 65 70 75 80 Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly Tyr Phe Thr Leu Leu Gly 85 90 95 Leu Pro Ala Met Leu Gln Ala Val Arg Ala Ala Ile Gln His Ser Gly 100 105 110 Gly Arg Ser Arg Arg Ala Arg Thr Lys Thr His Leu Arg Arg Gly Ser 115 120 125 Glu <210> 64 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Overlapping peptides within the first extracellular domain as disclosed in WO2008 / 145338 <400> 64 Met Asp Gln Trp Ser Thr Gln Asp Leu Tyr Asn Asn Pro Val Thr 1 5 10 15 <210> 65 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Overlapping peptides within the first extracellular domain as disclosed in WO2008 / 145338 <400> 65 Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly Leu 1 5 10 15 <210> 66 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Overlapping peptides within the first extracellular domain as disclosed in WO2008 / 145338 <400> 66 Val Phe Asn Tyr Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser 1 5 10 15 <210> 67 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Overlapping peptides within the first extracellular domain as disclosed in WO2008 / 145338 <400> 67 Gln Gly Leu Trp Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr 1 5 10 15 <210> 68 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Overlapping peptides within the first extracellular domain, as disclosed in WO2008 / 145338 <400> 68 Arg Ser Cys Val Arg Glu Ser Ser Gly Phe Thr Glu Cys Arg Gly 1 5 10 15 <210> 69 <211> 15 <212> PRT <213> Artificial sequence <220> <223> C-terminal epitope of CLDN18.2, as disclosed in WO2013 / 167259 <400> 69 Thr Glu Asp Glu Val Gln Ser Tyr Pro Ser Lys His Asp Tyr Val 1 5 10 15 <210> 70 <211> 12 <212> PRT <213> Artificial sequence <220> <223> C-terminal epitope of CLDN18.2, as disclosed in WO2013 / 167259 <400> 70 Glu Val Gln Ser Tyr Pro Ser Lys His Asp Tyr Val 1 5 10 <210> 71 <211> 15 <212> DNA <213> Artificial sequence <220> <223> cCl1-1, cCl1-2, hCl1a HCDR1 <400> 71 gactacgcga tgcac 15 <210> 72 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 HCDR2 <400> 72 tggatcaaca cgtacacggg gaagccgaca tacgcggacg acttcaaggg g 51 <210> 73 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1, cCl1-3 HCDR3 <400> 73 gccgtcttct acggatatac gatggacgcg 30 <210> 74 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 VH <400> 74 cagatccagc tcgtccagag cgggccggag ctgaagaagc cgggggagag cgtgaagatc 60 tcgtgcaagg cgagcggata tacgttcacg gactacgcga tgcactgggt caagcaagcg 120 ccggggaaag ggctgaagtg gatggggtgg atcaacacgt acacggggaa gccgacatac 180 gcggacgact tcaaggggcg attcgtgttc tcgctggagg cgagcgcgag cacggcgaac 240 ctgcaaatct cgaacctgaa gaacgaggac acggcgacgt acttctgcgc gcgggccgtc 300 ttctacggat atacgatgga cgcgtggggg cagggtacca gcgtgacggt ctcgagc 357 <210> 75 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 LCDR1 <400> 75 cgggcgagcg aggacatcta ctcgaacctg gcg 33 <210> 76 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 LCDR2 <400> 76 tccgtcaagc ggctgcaaga c 21 <210> 77 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 LCDR3 <400> 77 ctgcaaggga gcaacttccc gctgacg 27 <210> 78 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 VL <400> 78 gacatccaga tgacgcagag cccggcgtcg ctgagcgcga gcctggggga gacgatctcg 60 atcgcgtgcc gggcgagcga ggacatctac tcgaacctgg cgtggtatca acagaagagc 120 gggaagagcc cgcagctgct gatcttctcc gtcaagcggc tgcaagacgg cgtcccgagc 180 cgattctcgg ggagcgggag cgggacgcag tactcgctga agatctcggg gatgcagccg 240 gaggacgagg gggactactt ctgcctgcaa gggagcaact tcccgctgac gttcgggtcg 300 ggtaccaaac tcgagatcaa a 321 <210> 79 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> cCl1-2 HCDR2 <400> 79 tggatcaacg cgtacacggg gaagccgacc tacgcggacg acttcaaggg g 51 <210> 80 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> cCl1-2 HCDR3 <400> 80 gccgtctact acggatatac gatggac 27 <210> 81 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> cCl1-2 VH <400> 81 cagatccagc tcgtccagag cgggccggag ctgaagaagc cgggggagag cgtgaagatc 60 tcgtgcaaga cgagcggata tacgttcacg gactacgcga tgcactgggt caagcagggg 120 ccagggaaag ggatgaagtg gatggggtgg atcaacgcgt acacggggaa gccgacctac 180 gcggacgact tcaaggggcg attcgtgctg agcctggagg cgagcgcctc gacggcgaac 240 ctgcaaatct cgaacctgaa gaacgaggac acggcgacgt acttctgcgc gcgggccgtc 300 tactacggat atacgatgga cgcgtggggg cagggtacca gcgtgatcgt ctcgagc 357 <210> 82 <211> 33 <212> DNA <213> Artificial sequence <220> <223> cCl1-2 LCDR1 <400> 82 cggacgagcg aggacatcta ctcgaacttc gcg 33 <210> 83 <211> 21 <212> DNA <213> Artificial sequence <220> <223> cCl1-2 LCDR2 <400> 83 tcagtcaacc ggctgcaaga c 21 <210> 84 <211> 27 <212> DNA <213> Artificial sequence <220> <223> cCl1-2, cCl1-3, hCl1d, hCl1g LCDR3 <400> 84 ctgcaaggga gcaagttccc gctgacg 27 <210> 85 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> cCl1-1 VL <400> 85 gacatccaga tgacgcagag cccggcgagc ctgagcgcga gcctggggga gacgatctcg 60 atcgagtgcc ggacgagcga ggacatctac tcgaacttcg cgtggttcca gcagaagagc 120 gggaagagcc cgcagctgct gatctactca gtcaaccggc tgcaagacgg cgtcccgagc 180 cgattctcgg ggagcgggag cgggacgcag tactcgctga agatctcggg gatgcagccg 240 gaggacgagg gggactactt ctgcctgcaa gggagcaagt tcccgctgac gttcgggagc 300 ggtaccaaac tcgagatcaa a 321 <210> 86 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 HCDR1 <400> 86 gactacgcga tgtac 15 <210> 87 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 HCDR2 <400> 87 tggatcaaca cgtacacggg gaagccgacc tacgcggacg acttcaaggg g 51 <210> 88 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 VH <400> 88 cagatccagc tcgtccagag cgggccggag ctgaagaagc cgggggagag cgtgaagatc 60 tcgtgcaagg cgagcggata tacgttcacg gactacgcga tgtactgggt caagcaagtg 120 ccggggaaag ggctgcgatg gatggggtgg atcaacacgt acacggggaa gccgacctac 180 gcggacgact tcaaggggcg attcgtgttc tcgctggagg cgagcgcgag cacggcgaac 240 ctgcaaatct cgaacctgaa gaacgaggac acggcgacgt acttctgcgc gcgggccgtc 300 ttctacggat atacgatgga cgcgtggggg cagggtacca gcgtgacggt ctcgagc 357 <210> 89 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 LCDR1 <400> 89 cggacgagcg aggacatcta ctcgaacctg gcg 33 <210> 90 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 LCDR2 <400> 90 gcgatcaagc ggctgcaaga c 21 <210> 91 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> cCl1-3 VL <400> 91 gacatccaga tgacgcagag cccggcgagc ctgagcgcga gcctggggga gacgatctcg 60 atcgcgtgcc ggacgagcga ggacatctac tcgaacctgg cgtggtatca acagaagagc 120 gggaagagcc cgcagctgct gatcttcgcg atcaagcggc tgcaagacgg cgtcccgagc 180 cgattctcgg ggagcgggag cgggacgcag tactcgctga agatctcggg gatgcagccg 240 gaggacgagg gggactactt ctgcctgcaa gggagcaagt tcccgctgac gttcgggtcg 300 ggtaccaaac tcgagatcaa a 321 <210> 92 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> hCl1a HCDR2 <400> 92 tggatcaata catacacggg gaagccgact tatgcgcaaa aattccaagg a 51 <210> 93 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> hCl1a HCDR3 <400> 93 gcggtcttct acggatatac gatggatgcc 30 <210> 94 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> cCl1a VH <400> 94 caggtccaac tagtccaaag cggggcggaa gtcaagaagc ccggagcatc cgtcaaagtc 60 agctgcaagg cgagcggata tacatttacg gactacgcga tgcactgggt caggcaagcc 120 cctgggcaaa ggctcgaatg gatgggatgg atcaatacat acacggggaa gccgacttat 180 gcgcaaaaat tccaaggaag agtcacaatt acgcgggata catccgcatc taccgcctac 240 atggagctaa gctcgctgcg gagcgaggat acggcggtct actattgcgc ccgagcggtc 300 ttctacggat atacgatgga tgcctggggg cagggtaccc tggtcacggt ctcgagc 357 <210> 95 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> hCl1a, hCl1b, hCl1e, hCl1i LCDR1 <400> 95 agggcctccg aagacatcta ctccaacctg gca 33 <210> 96 <211> 21 <212> DNA <213> Artificial sequence <220> <223> hCl1a, hCl1b, hCl1e, hCl1i LCDR2 <400> 96 agcgtcaaaa gactacaaga t 21 <210> 97 <211> 27 <212> DNA <213> Artificial sequence <220> <223> hCl1a, hCl1b, hCl1e, hCl1i LCDR3 <400> 97 ttgcaaggaa gcaatttccc cttgact 27 <210> 98 <211> 321 <212> DNA <213> Artificial sequence <220> <223> cCl1a, hCl1b, hCl1e, hCl1i VL <400> 98 gacattcaaa tgacgcaaag cccatcatcg ctgagcgcat cggtcgggga tagagtcacc 60 ataacatgca gggcctccga agacatctac tccaacctgg catggtatca acaaaaaccg 120 gggaaggctc cgaagctgct gatatttagc gtcaaaagac tacaagatgg agtaccgagc 180 cgattttcgg gaagcgggag cgggacggat ttcacgctga ccatatcaag tttgcaaccg 240 gaggattttg cgacatacta ttgcttgcaa ggaagcaatt tccccttgac tttcgggcaa 300 ggtaccaagg tcgagatcaa a 321 <210> 99 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> hCl1b, hCl1c, hCl1d HCDR1 <400> 99 gattatgcaa tgcac 15 <210> 100 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> hCl1b, hCl1c, hCl1d HCDR2 <400> 100 tggattaaca cctacacggg caagcccaca tactcccaaa aattccaagg a 51 <210> 101 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> hCl1b, hCl1c, hCl1d HCDR3 <400> 101 gctgtattct atggatatac aatggatgcc 30 <210> 102 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> hCl1b, hCl1c, hCl1d VH <400> 102 caggtccaat tagtccaaag cggggcggaa gtcaagaagc cgggggcgag cgtcaaagtc 60 tcatgcaaag cgagcggata cacatttacg gattatgcaa tgcactgggt caggcaagca 120 cccggacaaa ggctggaatg gatgggatgg attaacacct acacgggcaa gcccacatac 180 tcccaaaaat tccaaggaag ggtcacgata acgagagaca cgagcgcgag caccggaatg 240 gatgggatgg attaacacct acacgggcaa gcccacatac tcccaaaaat tccaaggaag 300 ggtcacgata acgagagaca cgagcgcgag caccgtaccc tggtcaccgt ctcgagc 357 <210> 103 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> hCl1c, hCl1j LCDR1 <400> 103 cgaacgagcg aggacatata ctcaaacctt gca 33 <210> 104 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> hCl1c, hCl1j LCDR2 <400> 104 gcgataaaga ggctgcaaga c 21 <210> 105 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> hCl1c, hCl1j LCDR3 <400> 105 ttgcaaggct ccaaatttcc cctgaca 27 <210> 106 <211> 321 <212> DNA <213> Artificial sequence <220> <223> hCl1c, hCl1j VL <400> 106 gacatccaaa tgactcaaag cccatcatcg ctatcggcat cggtcgggga tagagtcacg 60 ataacatgcc gaacgagcga ggacatatac tcaaaccttg catggtatca acaaaagccg 120 gggaaggccc cgaagctact gatattcgcg ataaagaggc tgcaagacgg agttccatca 180 cgattttcgg gatctggctc ggggaccgat tttacgctga ctatatcatc gctgcaaccg 240 gaagattttg caacatacta ctgcttgcaa ggctccaaat ttcccctgac attcggacaa 300 ggtaccaagg tcgagatcaa a 321 <210> 107 <211> 33 <212> DNA <213> Artificial sequence <220> <223> hCl1d, hCl1g LCDR1 <400> 107 cggacgagcg aggatattta ttcgaacttt gca 33 <210> 108 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> hCl1d, hCl1g LCDR2 <400> 108 cagtcaatcg gctacaagat 20 <210> 109 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> hCl1d, hCl1g VL <400> 109 gacatccaaa tgacgcaatc accgagctcg ctgagcgcat ctgtcgggga ccgtgtcaca 60 atcacatgcc ggacgagcga ggatatttat tcgaactttg catggtatca acaaaaaccg 120 ggcaaggctc cgaaactttt gatttattca gtcaatcggc tacaagatgg cgtcccgagc 180 cgatttagcg ggagcggatc gggaaccgac tttacgctga cgatatcatc gctacaaccg 240 gaggacttcg cgacttatta ctgcctacaa gggagcaaat tcccgctgac attcggacaa 300 ggtaccaagg tcgagatcaa a 321 <210> 110 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> hCl1e HCDR1 <400> 110 gattacgcaa tgtac 15 <210> 111 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> hCl1e HCDR2 <400> 111 tggataaata cctatacggg aaagccaaca tacgcccaaa aattccaagg c 51 <210> 112 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> hCl1e HCDR3 <400> 112 gccgtctttt atggatatac gatggacgca 30 <210> 113 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> hCl1e VH <400> 113 caggtccaac tggtccaatc gggggctgaa gtcaaaaagc cgggggcgag cgtcaaagtc 60 agctgcaaag catcgggata cacatttacg gattacgcaa tgtactgggt caggcaagca 120 cccggccaac gactggaatg gatgggctgg ataaatacct atacgggaaa gccaacatac 180 gcccaaaaat tccaaggccg cgtcacaata acgcgggaca cgagcgcatc gacggcttat 240 atggaactat catcgctgcg atcggaagac acggcggtct attattgcgc acgcgccgtc 300 ttttatggat atacgatgga cgcatggggg cagggtaccc tggtcacggt ctcgagc 357 <210> 114 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1g HCDR1 <400> 114 gactacgcaa tgcac 15 <210> 115 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1g HCDR2 <400> 115 tggattaatg cctacacggg gaagccgacc tacgcacaaa aattccaagg a 51 <210> 116 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1g HCDR3 <400> 116 gccgtcttct atggatatac gatggatgct 30 <210> 117 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1g VH <400> 117 caggtccaat tggtccaaag cggggcggag gtcaagaagc cgggggcgag cgtcaaagtc 60 tcatgcaagg caagcggata tacatttacg gactacgcaa tgcactgggt ccggcaagcc 120 cctgggcaac ggctggaatg gatgggatgg attaatgcct acacggggaa gccgacctac 180 gcacaaaaat tccaaggacg agtcacgatt acgcgggata ctagcgcgag caccgcatat 240 atggagctaa gctcgctgcg atctgaggat accgctgtat actactgcgc gagagccgtc 300 ttctatggat atacgatgga tgcttggggg cagggtaccc tggtcacggt ctcgagc 357 <210> 118 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1h LCDR1 <400> 118 cgagcttcgg aggacatcta tagcaacttg gct 33 <210> 119 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1h LCDR2 <400> 119 agcgtcaaaa ggctccaaga c 21 <210> 120 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> hCl1f, hCl1h LCDR3 <400> 120 ctacaaggct ctaacttccc attgaca 27 <210> 121 <211> 321 <212> DNA <213> Artificial sequence <220> <223> hCl1f, hCl1h VL <400> 121 gatatccaaa tgacgcaatc accatctagc ctatcggcct ctgtggggga ccgagtcacc 60 atcacatgcc gagcttcgga ggacatctat agcaacttgg cttggtatca acaaaagccg 120 gggaaagcac caaagctgct gatatatagc gtcaaaaggc tccaagacgg agtcccaagc 180 cgattctcgg gctccggctc cgggacggat tttacgctga caatttcgag cctgcaaccg 240 gaggactttg caacctacta ttgcctacaa ggctctaact tcccattgac atttgggcaa 300 ggtaccaagg tcgagatcaa a 321 <210> 122 <211> 15 <212> DNA <213> Artificial sequence <220> <223> hCl1h, hCl1i, hCl1j HCDR1 <400> 122 gactacgcta tgtat 15 <210> 123 <211> 51 <212> DNA <213> Artificial sequence <220> <223> hCl1h, hCl1i, hCl1j HCDR2 <400> 123 tggattaatg cctacaccgg gaagccgact tatgcgcaaa aatttcaagg a 51 <210> 124 <211> 30 <212> DNA <213> Artificial sequence <220> <223> hCl1h, hCl1i, hCl1j HCDR3 <400> 124 gcggtctact atggatatac gatggacgca 30 <210> 125 <211> 357 <212> DNA <213> Artificial sequence <220> <223> hCl1h, hCl1i, hCl1j VH <400> 125 caggtccaac tggttcaatc tggagcggaa gtcaagaagc ccggagcatc cgtcaaagtc 60 tcgtgcaagg catctggata cacattcacc gactacgcta tgtattgggt ccggcaagcc 120 cccggacaac ggctggaatg gatgggatgg attaatgcct acaccgggaa gccgacttat 180 gcgcaaaaat ttcaaggaag ggtcacgatt acgcgggaca cgagcgcctc aaccgcatac 240 atggagctat cgagcctgcg aagcgaggac accgcggtct actactgcgc gcgggcggtc 300 tactatggat atacgatgga cgcatggggg cagggtaccc tggtcacggt ctcgagc 357 <210> 126 <211> 17 <212> PRT <213> Artificial sequence <220> <223> HC CDR2 is only for hCl1x, not for chimeric clones cCl1-1,2,3 <220> <221> Miscellaneous features <222> (4)..(4) <223> T or A <220> <221> Miscellaneous features <222> (12)..(12) <223> A or S <400> 126 Trp Ile Asn Xaa Tyr Thr Gly Lys Pro Thr Tyr Xaa Gln Lys Phe Gln 1 5 10 15 Gly <210> 127 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Constant light chain - CL domain <400> 127 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 128 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> Constant heavy chain - CH1+Fc domain <400> 128 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 129 <211> 330 <212> PRT <213> Synthetic Sequence <220> <223> L234A / L235A mutation in the constant heavy chain - CH1+Fc domain <400> 129 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 130 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> L236A / L236A / P329G mutation in the constant heavy chain - CH1+Fc domain <400> 130 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 131 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Sortase Tag <400> 131 Arg Leu Pro Gln Thr Gly Gly 1 5 <210> 132 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Sortase Tag <400> 132 Gly Gly Gly Gly Ser Leu Pro Gln Thr Gly Gly 1 5 10 <210> 133 <211> 261 <212> PRT <213> Homo sapiens <400> 133 Met Ala Val Thr Ala Cys Gln Gly Leu Gly Phe Val Val Ser Leu Ile 1 5 10 15 Gly Ile Ala Gly Ile Ile Ala Ala Thr Cys Met Asp Gln Trp Ser Thr 20 25 30 Gln Asp Leu Tyr Asn Asn Pro Val Thr Ala Val Phe Asn Tyr Gln Gly 35 40 45 Leucine Tryptophan Arginine Serine Cysteine Valine Arginine Glutamic acid Serine Serine Glycine Phenylalanine Threonine Glutamic acid Cysteine Arginine 50 55 60 Glycine Tyrosine Phenylalanine Threonine Leucine Leucine Glycine Leucine Proline Alanine Methionine Leucine Glutamine Alanine Valine Arginine 65 70 75 80 Alanine Leucine Methionine Isoleucine Valine Glycine Isoleucine Valine Leucine Glycine Alanine Isoleucine Glycine Leucine Leucine Valine 85 90 95 Serine Isoleucine Phenylalanine Alanine Leucine Lysine Cysteine Isoleucine Arginine Isoleucine Glycine Serine Methionine Glutamic acid Aspartic acid Serine 100 105 110 Alanine Lysine Alanine Asparagine Methionine Threonine Leucine Threonine Serine Glycine Isoleucine Methionine Phenylalanine Isoleucine Valine Serine 115 120 125 Glycine Leucine Cysteine Alanine Isoleucine Alanine Glycine Valine Serine Valine Phenylalanine Alanine Asparagine Methionine Leucine Valine 130 135 140 Threonine Asparagine Phenylalanine Tryptophan Methionine Serine Threonine Alanine Asparagine Methionine Tyrosine Threonine Glycine Methionine Glycine Glycine 145 150 155 160 Methionine Valine Glutamine Threonine Valine Glutamine Threonine Arginine Tyrosine Threonine Phenylalanine Glycine Alanine Alanine Leucine Phenylalanine 165 170 175 Valine Glycine Tryptophan Valine Alanine Glycine Glycine Leucine Threonine Leucine Isoleucine Glycine Glycine Valine Methionine Methionine 180 185 190 Cysteine Isoleucine Alanine Cysteine Arginine Glycine Leucine Alanine Proline Glutamic acid Glutamic acid Threonine Asparagine Tyrosine Lysine Alanine 195 200 205 Val Ser Tyr His Ala Ser Gly His Ser Val Ala Tyr Lys Pro Gly Gly 210 215 220 Phe Lys Ala Ser Thr Gly Phe Gly Ser Asn Thr Lys Asn Lys Lys Ile 225 230 235 240 Tyr Asp Gly Gly Ala Arg Thr Glu Asp Glu Val Gln Ser Tyr Pro Ser 245 250 255 Lys His Asp Tyr Val 260

Claims

1. An antibody or fragment thereof that binds to CLDN18.2, comprising: a. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:15, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively; b. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively; c. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:17, SEQ ID NO:14, and SEQ ID NO:11 respectively; d. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:16, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19, and SEQ ID NO:11 respectively; e. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:12, SEQ ID NO:15, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively; f. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6 respectively; g. HCDR1, HCDR2, and HCDR3 sequences that are SEQ ID NO:1, SEQ ID NO:20, and SEQ ID NO:3 respectively, and LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NO:18, SEQ ID NO:19, and SEQ ID NO:11 respectively; h. The HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively; or i. The HCDR1, HCDR2 and HCDR3 sequences of SEQ ID NO:12, SEQ ID NO:20 and SEQ ID NO:8 respectively, and the LCDR1, LCDR2 and LCDR3 sequences of SEQ ID NO:17, SEQ ID NO:14 and SEQ ID NO:11 respectively.

2. The antibody or fragment thereof of claim 1, comprising: a. The VH sequence of SEQ ID NO:33 and the VL sequence of SEQ ID NO:38; b. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:38; c. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:39; d. The VH sequence of SEQ ID NO:34 and the VL sequence of SEQ ID NO:40; e. The VH sequence of SEQ ID NO:35 and the VL sequence of SEQ ID NO:38; f. The VH sequence of SEQ ID NO:36 and the VL sequence of SEQ ID NO:41; g. The VH sequence of SEQ ID NO:36 and the VL sequence of SEQ ID NO:40; h. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:41; i. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:38; or j. The VH sequence of SEQ ID NO:37 and the VL sequence of SEQ ID NO:

39.

3. The antibody or fragment thereof of claim 1, comprising: a. The heavy chain sequence of SEQ ID NO:46 and the light chain sequence of SEQ ID NO:51; b. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:51; c. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:52; d. The heavy chain sequence of SEQ ID NO:47 and the light chain sequence of SEQ ID NO:53; e. The heavy chain sequence of SEQ ID NO:48 and the light chain sequence of SEQ ID NO:51; f. The heavy chain sequence of SEQ ID NO:49 and the light chain sequence of SEQ ID NO:54; g. The heavy chain sequence of SEQ ID NO:49 and the light chain sequence of SEQ ID NO:53; h. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:54; i. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:51; j. The heavy chain sequence of SEQ ID NO:50 and the light chain sequence of SEQ ID NO:52, or an engineered Fc domain version thereof.

4. An antibody or fragment thereof that binds to CLDN18.2 and comprises: a. The HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, respectively; b. The HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:1, SEQ ID NO:7, and SEQ ID NO:8, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, respectively; or c. The HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:12, SEQ ID NO:2, and SEQ ID NO:3, respectively, and the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:11, respectively.

5. The antibody or fragment thereof of claim 4, which comprises: a. The VH sequence of SEQ ID NO:27 and the VL sequence of SEQ ID NO:28; b. The VH sequence of SEQ ID NO:29 and the VL sequence of SEQ ID NO:30; or c. The VH sequence of SEQ ID NO:31 and the VL sequence of SEQ ID NO:

32.

6. The antibody or fragment thereof of any one of claims 1 to 5, wherein the antibody or fragment thereof a. is IgA1, IgA2, IgD, IgE, IgG1, IgG2, IgG3, IgG4, synthetic IgG, IgM, F(ab)2, scFv, IgGACH2, F(ab’)2, scFvCH3, Fab, scFv4, scFv3, scFv2, dsFv, Fv, scFv-Fc, (scFv)2, non-consumable IgG, bispecific antibody, bivalent antibody, or an Fc-engineered version thereof; b. is humanized; c. does not bind to CLDN18.1; d. is isolated; and / or e. has a lower susceptibility to post-translational deamidation compared to a control antibody, wherein the control antibody comprises the heavy chain sequence of SEQ ID NO:55 and the light chain sequence of SEQ ID NO:

56.

7. The antibody or fragment thereof of any one of claims 1 to 5, wherein the binding to tumor tissue expressing CLDN18.2 is increased compared to healthy tissue expressing CLDN18.2, as measured by flow cytometry or by immunohistochemistry.

8. An antibody or fragment thereof according to any one of claims 1 to 5, wherein the antibody or fragment thereof binds to CLDN18.2 expressed in HEK293T cells or PA-TU-8988-High cells with an EC50 value that is at least 1.1-fold but not more than 3-fold the EC50 value of a control antibody binding to CLDN18.2 expressed in HEK293T cells or PA-TU-8988-High cells, the control antibody comprising the heavy chain sequence of SEQ ID NO:55 and the light chain sequence of SEQ ID NO:56, wherein binding is measured by flow cytometry (FC) titration.

9. A nucleic acid encoding an antibody or fragment thereof according to any one of claims 1 to 8.

10. A vector comprising the nucleic acid of claim 9.

11. A host cell comprising the nucleic acid of claim 9 or the vector of claim 10.

12. Use of an antibody or fragment thereof according to any one of claims 1 to 8, the nucleic acid of claim 9, the vector of claim 10 or the host cell of claim 11 in the preparation of a medicament for treating a subject suffering from a neoplastic disease; wherein the neoplastic disease is selected from pancreatic cancer and gastric cancer.

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