Patent Publication Number: US-2022235128-A1

Title: Anti-cldn antibody and pharmaceutical composition thereof and detection method therefor

Description:
TECHNICAL FIELD 
     The present invention relates to an anti-CLDN antibody, the present invention also relates to a pharmaceutical composition comprising the anti-CLDN antibody, and the present invention also relates to a method for detecting whether CLDN is present in a biological sample. 
     BACKGROUND 
     Tight junction (TJ) plays a key role in the material flow between cells, maintains cell polarity by blocking the radial diffusion of membrane proteins and membrane lipids. In addition, it also participates in recruiting signal molecules that regulate cell proliferation, differentiation and movement. Tight junction is formed by claudin (CLDN), and the claudin family consists of more than 20 protein molecules, all of which contain a quadruple transmembrane domain and similar amino acid sequences, but their tissue distribution is specific. Human CLDN genes are distributed in pairs on different chromosomes, which implies that some CLDN genes are derived from gene replication. 
     The molecular weight of CLDN protein is mostly in the range of 20-34 kDa, and the biggest difference is the sequence and size of intracellular C-terminal, which contains a PDZ domain binding motif, which enables CLDN protein to directly interact with tight junction related proteins in cytoplasm, such as ZO-1, ZO-2, ZO-3 and MUPP1. In addition, this sequence contains post-transcriptional modification sites such as phosphorylation sites, which can affect the localization and function of protein molecules. MAPK (Mitogen-activated protein kinase) or PKC (protein kinase C) can phosphorylate CLDN1, and cAMP (cyclic AMP) can induce the phosphorylation of CLDN5, all of which can promote the barrier function of CLDN protein, while PKA-mediated phosphorylation of CLDN16 can enhance magnesium ion transport. 
     CLDN plays a key role in regulating selective permeation of cell bypass. CLDN2 and CLDN15 participate in the formation of cation channels and cation pores, while CLDN4/7/10 participate in the formation of anion channels and pores. Claudin protein is highly expressed in some cell lines, which affects transepithelial electrical resistance and permeability. In cultured epidermal-derived cells, CLDN1/4/5/7 can increase transepithelial electrical resistance, while CLDN2 and CLDN10 do not. 
     CLDN gene mutation is deemed relevant to various diseases. CLDN1 mutation may lead to sclerosing cholangitis and ichthyosis, while CLDN16 and CLDN19 mutations are considered to be related to hypomagnesemia and hypercalcinuria. 
     Differential expression of CLDN protein is thought to be associated with various cancers. CLDN1 and CLDN7 are down-regulated in invasive breast cancer, prostate cancer and esophageal cancer, while CLDN3/4 are found to be up-regulated to varying degrees in cervical cancer, colon cancer, esophageal cancer, gastric cancer and other cancers. Sahin et al. found that in normal tissues, the isoform 2 subtype of CLDN18 (CLDN18.2) was only expressed in the differentiated epidermal cells of the gastric mucosa, but not in the area of gastric stem cells, while abnormally high expression was found in primary gastric cancer and its metastases. High expression of CLDN18.2 has also been reported in pancreatic cancer, esophageal cancer and lung cancer. Because CLDN18.2 is located on the surface of cell membrane, its biological function and characteristics determine that it is an ideal therapeutic target. In recent years, monoclonal antibodies against this target have also appeared, and among them, IMAB362 (Claudiximab) of Ganymed Company is the fastest developing one. IMAB362 binds CLDN18.2 on the surface of tumor cells, inducing antibody-dependent cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), thus killing tumor cells. When combined with chemotherapy, IMAB362 can also enhance T cell infiltration and up-regulate pro-inflammatory factors. 
     SUMMARY OF THE INVENTION 
     The purpose of the present invention is to provide an anti-CLDN antibody, a pharmaceutical composition thereof and a detection method therefor. 
     The present invention adopts the following technical solutions. 
     An anti-CLDN antibody which comprises a heavy chain and a light chain: 
     wherein, the heavy chain of the antibody contains one or more CDRs, and the CDR of the heavy chain is no more than three amino acids different from the CDR sequence of any one of SEQ ID No: 1 to SEQ ID No: 7 or any one of SEQ ID No: 15 to SEQ ID No: 30, 
     the light chain of the antibody contains one or more CDRs, and the CDR of the light chain is no more than three amino acids different from the CDR sequence of any one of SEQ ID No: 8 to SEQ ID No: 14 or any one of SEQ ID No: 31 to SEQ ID No: 46. 
     Further, the anti-CLDN antibody of the present invention has a feature that the heavy chain of the antibody is selected from any one of SEQ ID No: 1 to No. 7. 
     Further, the anti-CLDN antibody of the present invention has a feature that the light chain of the antibody is selected from any one of SEQ ID No: 8 to SEQ ID No: 14 or any one of SEQ ID No: 31 to SEQ ID No: 46. 
     Further, the anti-CLDN antibody of the present invention has a feature that the heavy chain of the antibody is selected from any one of SEQ ID No: 15 to SEQ ID No: 30. 
     Further, the anti-CLDN antibody of the present invention has a feature that the light chain of the antibody is selected from any one of SEQ ID No: 31 to SEQ ID No: 46. 
     Further, the anti-CLDN antibody of the present invention has a feature that the heavy chain and the light chain of the antibody form a combination selected from the group consisting of: 
     SEQ ID No: 1 and SEQ ID No: 8, SEQ ID No: 2 and SEQ ID No: 9, SEQ ID No: 3 and SEQ ID No: 10, 
     SEQ ID No: 4 and SEQ ID No: 11, SEQ ID No: 5 and SEQ ID No: 12, SEQ ID No: 6 and SEQ ID No: 13, 
     SEQ ID No: 7 and SEQ ID No: 14, SEQ ID No: 15 and SEQ ID No: 31, SEQ ID No: 16 and SEQ ID No: 32, 
     SEQ ID No: 17 and SEQ ID No: 33, SEQ ID No: 18 and SEQ ID No: 34, SEQ ID No: 19 and SEQ ID No: 35, 
     SEQ ID No: 20 and SEQ ID No: 36, SEQ ID No: 21 and SEQ ID No: 37, SEQ ID No: 22 and SEQ ID No: 38, 
     SEQ ID No: 23 and SEQ ID No: 39, SEQ ID No: 24 and SEQ ID No: 40, SEQ ID No: 25 and SEQ ID No: 41, 
     SEQ ID No: 26 and SEQ ID No: 42, SEQ ID No: 27 and SEQ ID No: 43, SEQ ID No: 28 and SEQ ID No: 44, 
     SEQ ID No: 29 and SEQ ID No: 45, SEQ ID No: 30 and SEQ ID No: 46. 
     The present invention also provides a polynucleotide encoding the antibody as defined hereinabove. 
     The present invention also provides a pharmaceutical composition comprising any one of the above antibodies. 
     The present invention also provides the application of any one of the above-mentioned antibodies in the preparation of anti-tumor drugs. 
     The present invention also provides a method for detecting the presence or absence of CLDN in a biological sample, which comprises the steps of administering an antibody as described in any one of the above to the biological sample, wherein the antibody has a detectable label, and the steps of detecting the presence or absence of the detectable label or detecting the content of the detectable label. 
     Beneficial Effects of the Invention 
     The anti-CLDN antibody of the present invention has stronger binding ability with cells than IMAB362. Moreover, the antibody of the present invention shows better effect of inhibiting tumor growth than IMAB362 in animal pharmacodynamics in vivo. 
    
    
     
       DESCRIPTION OF DRAWINGS 
         FIG. 1A  shows the binding ability of control antibody QP024025 to CHOS cells 
         FIG. 1B  shows the binding ability of antibody QP188189 screened out from hybridoma to CHOS cells 
         FIG. 1C  shows the binding ability of antibody QP190191 screened out from hybridoma to CHOS cells 
         FIG. 1D  shows the binding ability of antibody QP192193 screened out from hybridoma to CHOS cells 
         FIG. 1E  shows the binding ability of antibody QP196198 screened out from hybridoma to CHOS cells 
         FIG. 1F  shows the binding ability of antibody QP199200 screened out from hybridoma to CHOS cells 
         FIG. 1G  shows the binding ability of antibody QP201202 screened out from hybridoma to CHOS cells 
         FIG. 1H  shows the binding ability of antibody QP207208 screened out from hybridoma to CHOS cells  FIG. 2A  shows the binding ability of antibody QP10731074 screened out from phage to CHOS cells. 
         FIG. 2B  shows the binding ability of antibody QP10791080 screened out from phage to CHOS cells. 
         FIG. 2C  shows the binding ability of antibody QP10851086 screened out from phage to CHOS cells. 
         FIG. 2D  shows the binding ability of antibody QP10911092 screened out from phage to CHOS cells. 
         FIG. 2E  shows the binding ability of antibody QP10971098 screened out from phage to CHOS cells. 
         FIG. 2F  shows the binding ability of antibody QP10991100 screened out from phage to CHOS cells. 
         FIG. 3A  shows the binding ability of antibody QP11051106 screened out from phage to CHOS cells. 
         FIG. 3B  shows the binding ability of antibody QP11071108 screened out from phage to CHOS cells. 
         FIG. 3C  shows the binding ability of antibody QP11091110 screened out from phage to CHOS cells. 
         FIG. 3D  shows the binding ability of antibody QP11111112 screened out from phage to CHOS cells. 
         FIG. 3E  shows the binding ability of antibody QP11131114 screened out from phage to CHOS cells. 
         FIG. 3F  shows the binding ability of antibody QP11151116 screened out from phage to CHOS cells. 
         FIG. 4A  shows the binding ability of antibody QP11171118 screened out from phage to CHOS cells. 
         FIG. 4B  shows the binding ability of antibody QP11031104 screened out from phage to CHOS cells. 
         FIG. 4C  shows the binding ability of antibody QP10451046 screened out from phage to CHOS cells. 
         FIG. 4D  shows the binding ability of antibody QP10471048 screened out from phage to CHOS cells. 
         FIG. 5A  shows the binding ability of control antibody QP024025 to different 293T transient cell lines. 
         FIG. 5B  shows the binding ability of antibody QP188189 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5C  shows the binding ability of antibody QP190191 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5D  shows the binding ability of antibody QP192193 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5E  shows the binding ability of antibody QP196198 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5F  shows the binding ability of antibody QP199200 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5G  shows the binding ability of antibody QP201202 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 5H  shows the binding ability of antibody QP207208 screened out from hybridoma to different 293T transient cell lines. 
         FIG. 6A  shows the binding ability of antibody QP10451046 screened out from phage to different 293T transient cell lines. 
         FIG. 6B  shows the binding ability of antibody QP10711072 screened out from phage to different 293T transient cell lines. 
         FIG. 6C  shows the binding ability of antibody QP10731074 screened out from phage to different 293T transient cell lines. 
         FIG. 6D  shows the binding ability of antibody QP10851086 screened out from phage to different 293T transient cell lines. 
         FIG. 6E  shows the binding ability of antibody QP10911092 screened out from phage to different 293T transient cell lines. 
         FIG. 6F  shows the binding ability of antibody QP10991100 screened out from phage to different 293T transient cell lines. 
         FIG. 6G  shows the binding ability of antibody QP11031104 screened out from phage to different 293T transient cell lines. 
         FIG. 6H  shows the binding ability of antibody QP11051106 screened out from phage to different 293T transient cell lines. 
         FIG. 7A  shows the binding ability of antibody QP11071108 screened out from phage to different 293T transient cell lines. 
         FIG. 7B  shows the binding ability of antibody QP11091110 screened out from phage to different 293T transient cell lines. 
         FIG. 7C  shows the binding ability of antibody QP11111112 screened out from phage to different 293T transient cell lines. 
         FIG. 7D  shows the binding ability of antibody QP11131114 screened out from phage to different 293T transient cell lines. 
         FIG. 7E  shows the binding ability of antibody QP11151116 screened out from phage to different 293T transient cell lines. 
         FIG. 7F  shows the binding ability of antibody QP11171118 screened out from phage to different 293T transient cell lines. 
         FIG. 8A  is the curve of control group IMAB362 in the experiment of binding ability of control antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8B  is the curve of QP190191 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8C  is the curve of QP192193 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8D  is the curve of QP201202 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8E  is the curve of QP207208 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8F  is the curve of QP11091110 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8G  is the curve of QP11131114 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 8H  is the curve of QP11151116 in the experiment of binding ability of antibody to gastric cancer PDX model GA0006 tumor cells. 
         FIG. 9  shows the pharmacodynamic test of the antibody against the gastric cancer PDX model GA0006. 
         FIG. 10A  is the tumor growth curve of PBS group (negative control) after grouping. 
         FIG. 10B  is the tumor growth curve of QP192193 group after grouping. 
         FIG. 10C  is the tumor growth curve of QP207208 group after grouping. 
         FIG. 10D  is the tumor growth curve of QP11151116 group after grouping. 
         FIG. 10E  is the tumor growth curve of the control antibody IMAB362 group after grouping. 
         FIG. 11  shows the weight of tumor at D31 in each group of mice added with each antibody of the present invention and a control antibody. 
         FIG. 12  is a real shot diagram of the tumor volume of each experimental group. 
         FIG. 13  is the body weight curve of mice in each group. 
         FIG. 14  is the curve of body weight change rate of mice in each group. 
     
    
    
     DETAILED DESCRIPTION 
     Specific embodiments of the present invention will be explained below with reference to the accompanying drawings. 
     Antibodies of the present invention include, but are not limited to, human antibodies. In various antibody screening methods provided by the present invention, the more convenient hybridoma screening antibody is usually preferred. However, the antigen of this target (claudin 18.2) is difficult to obtain, so the phage library is also used to screen the antibody. The sequences of the anti-claudin18.2 antibodies screened out in this embodiment are as follows: 
     For Antibodies Screened by Hybridoma, the CDRs in the Heavy Chain Variable Region (VH) are Shown in the Underlined Parts of the Sequences 
     
       
         
           
               
            
               
                 &gt; QP189 
               
               
                 QVQLQQSGAELVKPGASVKLSCKASGYTFT SYGIN WVRQRPEQGLEWIG WL   
               
               
                   
               
               
                   FPGDGTIKYNENFKG KATLTTDRSSSAAYMQLSRLTSEDSAVYFCAR GGYY   
               
               
                   
               
               
                   GNAMDY WGQGTSVTVSS 
               
               
                   
               
               
                 &gt; QP191 
               
               
                 EVKLVESGGGLVKPGGSLKLSCAASGFTFS NYAMS WVRQTPEKRLEWVA SI   
               
               
                   
               
               
                   ISGGRTYYLDSEKG RFTISRDNARNNLYLQMSSLRSEDTAMYYCTR IYYGN   
               
               
                   
               
               
                   SFDY WGQGTTLTVSS 
               
               
                   
               
               
                 &gt; QD193 
               
               
                 QVQLQQSGAELVRPGSSVKISCKASGYAFS SYWMN WVKQRPGQGLEWIG QI   
               
               
                   
               
               
                   YPGNGDTTYNGKFKG QATLTADKSSSTVYMQLSSLTSEDSAVYFCAR FVKG   
               
               
                   
               
               
                   NAMDY WGQGTSVTVSS 
               
               
                   
               
               
                 &gt; QD198 
               
               
                 QVQLKESGPGLVAPSQSLSITCTVSGFSLT IYGVH WVRQPPGRGLEWLG VI   
               
               
                   
               
               
                   WAGGSTNYNSALMS RLSISKDNSKSQVFLKVNSLQTDDTAMYYCAR DYYYG   
               
               
                   
               
               
                   SGFDY WGQGTTLTVSS 
               
               
                   
               
               
                 &gt; QD200 
               
               
                 DVQLVESGGGLVQPGGSRKLSCAASGFTFS SFGMH WVRQAPEKGLEWVA YI   
               
               
                   
               
               
                   SSGSNSIYYVDTVKG RFTISRDNPKNTLFLQMTSLKSEDTAMYYCAR NAYY   
               
               
                   
               
               
                   GNSFDY WGQGTTLTVSS 
               
               
                   
               
               
                 &gt; QD202 
               
               
                 EVQLQQSGPELVKPGASVKMSCKASGYTFT NYFVH WVKQKPGQGLEWIG YI   
               
               
                   
               
               
                   NPYNDDTKYNEKFKG KATLTSDKSSSTAYMDLSSLTSEDSAVYYC LSLRFF   
               
               
                   
               
               
                   AY WGQGTLVTVSA 
               
               
                   
               
               
                 &gt; QD208 
               
               
                 EVQLQQSGPELVKPGASVKMSCKASGYTFT SYIMH WVKQKPGQGLEWIG YI   
               
               
                   
               
               
                   NPYNDGTKYNEKFKG KATLTSDKSSSTVYMELSSLTSEDSAVYCCAR LGFT   
               
               
                   
               
               
                   TRNAMDY WGQGTSVTVSS 
               
            
           
         
       
     
     Light Chain Sequence: 
     The CDRs in the light chain variable region (VL) are shown in the underlined parts of the sequences. 
     
       
         
           
               
            
               
                 &gt; QD188 
               
               
                 DIVMTQSPSSLTVTAGEKVTMSC KSSQSLLNSGNQKSYLT WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY WASTRES GVPDRFTGSGSGTDFTLTISSVQAEDLAVYFC QNDYFYPY   
               
               
                   
               
               
                   T FGGGTKLEIK 
               
               
                   
               
               
                 &gt; QD190 
               
               
                 DIVMTQSPSSQTVTAGEKVTMSC KSSQSLLNSGNQKNYLT WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY WASTRES GVPDRFTGSGSGTDFTLTISNMQAEDLAVYYC QNDYSYPF   
               
               
                   
               
               
                   T FGSGTKLEIK 
               
               
                   
               
               
                 &gt; QD192 
               
               
                 DIVMTQSPSSLTVTAGEKVTMSC KSSQSLLNSGNQKNYLT WYQQNPGQPPK 
               
               
                   
               
               
                 MLIY WASTRES GVPDRFTGSGSGIDFSLTISSVQAEDLALYYC QNAYSYPF   
               
               
                   
               
               
                   T FGSGTKLEIK 
               
               
                   
               
               
                 &gt; QD196 
               
               
                 DIVMTQSPSSLSVSAGEKVTMSC KSSQSLLNSGNQKNYLA WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY GASTRES GVPDRFTGSGSGTDFTLTISSVRAEDLAVYYC QNDHYYPF   
               
               
                   
               
               
                   T FGSGTKLEIK 
               
               
                   
               
               
                 &gt; QD199 
               
               
                 DIVMTQSPSSLTVTAGEKVTMSC KSSQSLLNSGNQKNYLT WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY WASTRES GVPDRFTGSGSGTVFTLTISSVQAEDLAVYFC QNNYYYPL   
               
               
                   
               
               
                   T FGAGTKLELK 
               
               
                   
               
               
                 &gt; QD201 
               
               
                 DIVMTQSPSSLTVTAGEKVTMSC KSSQSLLNSGNQKNYLT WYQQKPGQAPK 
               
               
                   
               
               
                 LLIY WASTRES GVPDRFIGSGSGTDFTLTISHVQAEDLAVYFC QNDYSYPL   
               
               
                   
               
               
                   T FGAGTNLELK 
               
               
                   
               
               
                 &gt; QD207 
               
               
                 DIVMTQSPSSLSVSAGEKVTMNC KSSQSLLNSGNQKNYLA WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY GASTRESG VPDRFTGSGSGTDFTLTISSVQAEDLAVYYC QNDHSYP F 
               
               
                   
               
               
                 TFGSGTKLEIK 
               
            
           
         
       
     
     For Antibodies Screened by Phage Library, the CDRs in the Heavy Chain Variable Region (VH) are Shown in the Underlined Parts of the Sequences 
     
       
         
           
               
            
               
                 &gt; QD1045 
               
               
                 QVQLVQSGAEVKKPGASVKVSCKASGYTFT SYYMH WVRQAPGQGLEWMG II   
               
               
                   
               
               
                   NPSGGSTSYAQKFQG RVTMTRDTSTSTVYMELSSLRSEDTAVYYCAR DYAF   
               
               
                   
               
               
                   TGFDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1047 
               
               
                 QVQLVQSGAEVKKPGASVKVSCKASGYTFT SYYMH WVRQAPGQGLEWMG II   
               
               
                   
               
               
                   NPSGGSTSYAQKFQG RVTMTRDTSTSTVYMELSSLRSEDTAVYYCAR SSAY   
               
               
                   
               
               
                   GTYSMDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1073 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTF SSYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTGVYYCAR GSGS   
               
               
                   
               
               
                   WFGPYFDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1079 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR TDGA   
               
               
                   
               
               
                 TPFDYWGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1085 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR RSYY   
               
               
                   
               
               
                   GTGAFDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1091 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR SLGY   
               
               
                   
               
               
                   FSGLAFDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1097 
               
               
                   
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR GYNW   
               
               
                   
               
               
                   SFGMDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1099 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR AGYF   
               
               
                   
               
               
                   PRSLDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1103 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR GYSW   
               
               
                   
               
               
                   YWLFGFDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1105 
               
               
                 QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS WVRQAPGQGLEWMG GI   
               
               
                   
               
               
                   IPIFGTANYAQKFQG RVTITADKSTSTAYMELSSLRSEDTAVYYCAR GGDW   
               
               
                   
               
               
                   GGYMDY WGQGTTVTVSS 
               
               
                   
               
               
                 &gt; QD1107 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR DYYY   
               
               
                   
               
               
                   YFWFDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1109 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR DYYY   
               
               
                   
               
               
                   YYWFDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1111 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GYYY   
               
               
                   
               
               
                   YFWFDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1113 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK SAAY   
               
               
                   
               
               
                   YYFWFDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1115 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR DSYY   
               
               
                   
               
               
                   YYFWYDY WGQGTLVTVSS 
               
               
                   
               
               
                 &gt; QD1117 
               
               
                 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMS WVRQAPGKGLEWVS AI   
               
               
                   
               
               
                   SGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK AIDY   
               
               
                   
               
               
                   YTFDY WGQGTLVTVSS 
               
            
           
         
       
     
     Light Chain Sequence: 
     The CDRs in the Light Chain Variable Region (VL) are Shown in the Underlined Parts of the Sequences. 
     
       
         
           
               
            
               
                 &gt; QD1046 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQALMTPT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1048 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQDLWPRT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1074 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQAAQSPT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1080 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQALNTPPT   
               
               
                   
               
               
                 FGQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1086 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVTDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQALMTPT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1092 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQGRQFPT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1098 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQGLNTFT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1100 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQALQWDT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1104 
               
               
                 DIVMTQSPLSLPVTPGEPASISC RSSQSLLHSNGYNYLD WYLQKPGQSPQL 
               
               
                   
               
               
                 LIY LGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYC MQALQTGT F 
               
               
                   
               
               
                 GQGTKVEIK 
               
               
                   
               
               
                 &gt; QD1106 
               
               
                 DIVMTQSPDSLAVSLGERATINC KSSQSVLYSSNNKNYLA WYQQKPGQPPK 
               
               
                   
               
               
                 LLIY WASTRES GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC QQYYTTPF   
               
               
                   
               
               
                   T FGQGTKVE 
               
               
                   
               
               
                 IK 
               
               
                   
               
               
                 &gt; QD1108 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYNSYST FGQGTK 
               
               
                   
               
               
                 VEIK 
               
               
                   
               
               
                 &gt; QD1110 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYSSYSPT FGQGT 
               
               
                   
               
               
                 KVEIK 
               
               
                   
               
               
                 &gt; QD1112 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTKFTLTISSLQPDDFATYYC QQYSTYPLT FGQGT 
               
               
                   
               
               
                 KVEIK 
               
               
                   
               
               
                 &gt; QD1114 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYLSYPPT FGQGT 
               
               
                   
               
               
                 KVEIK 
               
               
                   
               
               
                 &gt; QD1116 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYNSYPLT FGQGT 
               
               
                   
               
               
                 KVEIK 
               
               
                   
               
               
                 &gt; QD1118 
               
               
                 DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DA   
               
               
                   
               
               
                   SSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYSTYPLT FGQGT 
               
               
                   
               
               
                 KVEVK 
               
            
           
         
       
     
     Antibody Binding Ability Experiment: 
     Experiment 1: Detection of the binding ability of antibody to tumor cell line by FACS 
     a) 96-well plate was planted with 2×10 5  cells per well, centrifuged at 1000×rpm for 5 minutes. Cells were washed once with 1×PBS, and excess liquid of supernatant was sucked out; 
     b) 3% BSA-PBS solution was added to block the cells and the cells were blocked at 4° C. for 60 minutes. 
     c) The antibody to be tested was diluted to 5 ug/ml with blocking solution, added into the wells, and incubated at 4° C. for 60 minutes. 
     d) The antibody was sucked out, and cells in each well were washed with 220 ul of 1×PBS for three times. 
     e) 50 ul of PE-anti-human FC (1:200 dilution) secondary antibody was added to each well, and incubated in the dark at 4° C. for 40 minutes. 
     f) The antibody was sucked out, and cells in each well were washed with 1×PBS for four times. 
     g) On-board detection was performed. 
     Experiment 2: Detection of the binding ability of antibody to PDX tissue tumor cell by FACS 
     a) A gentleMACS™ C Tube and 3 ml of digestion buffer were prepared for each tumor. The digestion buffer was prepared according to the instructions of Tumor Dissemination Kit (miltenyibiotech, 130-096-730) and it was freshly prepared just before use. 
     b) The mice were sacrificed, the tumor was removed with cleaning tools, and the blood vessels, fat, fascia and other tissues attached to the tumor surface were removed after washing with 1×PBS. Each digestion tube could digest no more than 0.8 g of tumor tissue to ensure that the tumor tissue was completely digested by digestion buffer. 
     c) The tumor tissue was placed in the hole of 6-well plate, digestion buffer was added, and the tumor tissue was cut to small pieces of about 1 mm 3  with clean tweezers and scalpel. 
     d) The tissue pieces were put into a gentleMACS™ C Tube, the well plate was washed with residual digestion buffer, and it was transferred into the digestion tube together, and placed on the ice. 
     e) The digestion tube was put upside down into gentleMACS automatic tissue processor, and the program 37_c_m_TDK_1 was selected for digestion. After the program was over, the digestion tube was removed and centrifuged instantaneously at 300×g to collect the cells and remaining tissues. 
     f) The supernatant was discarded, and the cells and tissues were re-suspended with 1×PBS. The cell suspension was added to the cell strainer, which was placed in a 50 ml centrifuge tube, and the suspension was sifted with 10 ml PBS to become single cell suspension. 
     g) After centrifuged at 300×g for 5 minutes, the supernatant was removed, and 5 ml PBS was used to re-suspend. Cell counting was performed and the cell concentration was adjusted to 1×10 6  cells per sample. 
     h) 100 ul of PBS solution with a concentration of 1 μg/ml Mouse BD Fc Block (CAT #553141) was added to each sample to re-suspend cells, 20 ul of human FcR Blocking Reagent was added, mixed well, and incubated in the dark at room temperature for 10 minutes. 
     i) 5 ug/ml of primary antibody was added and incubated in the dark at 4° C. for 60 minutes. 
     j) 2 ml of FACS wash buffer was added, and the cells were gently resuspended, centrifuged at 300×g for 5 minutes, and the supernatant was removed, that was repeated twice. 
     k) 100 ul of FACS wash buffer containing PE labeled human IgG Fc secondary antibody and dye was added to incubate cells in the dark for 60 minutes. 
     l) 2 ml of FACS wash buffer was added, and the cells were gently resuspended, centrifuged at 300×g for 5 minutes, and the supernatant was removed, that was repeated twice. 
     m) 200 ul of FACS wash buffer was used to resuspend the cells, and on-board detection was performed. 
     The hybridoma clone in CHOS system and phage clone in CHOS system in the experimental drawings refer to the binding ability of antibodies screened from hybridoma and phage to CHOS cells, respectively. In the figures, CHO18.2:CHOS cells transfected with the CLDN18.2 gene for establishment of stable CHOS-CLDN18.2 cells, CHO18.1: CHOS cells transfected with the CLDN18.1 gene for establishment of stable CHOS-CLDN18.1 cells, CHOS:CHOS cells transfected with empty vector. 
     CHOS is a cell line obtained by immortalization of hamster ovary cells. FACS (Flow Cytometry Fluorescence Sorting Technique) was used to detect the binding ability of antibody to the cell line. 
       FIGS. 1A to 1H  show the binding ability of antibodies screened by hybridoma to CHOS cells. 
       FIGS. 2A to 2H, 3A to 3H, and 4A to 4D  show the binding ability of phage-screened antibodies to CHOS cells. 
     Purpose: To verify the binding ability of antibody to claudin18.1 (nonspecific binding) and claudin18.2 in CHOS cell lines. 
     In the figures of CHOS cell lines, each panel is composed of FACS detection results of three CHOS cell lines with one antibody to be tested, wherein: 
     1. The red curve (3) indicates the binding ability of the antibody to be tested to the CHOS cell line (CHOS) transfected with the empty vector, that is, the negative control; 
     2. The blue curve (2) indicates the binding ability of the antibody to be tested to the CHOS cell line transfected with claudin 18.1 (CHO18.1), that is, the detection of non-specific binding. 
     3. The yellow curve (1) indicates the binding ability of the antibody to be tested to the CHOS cell line transfected with claudin 18.2 (CHO 18.2), that is, the detection of binding ability of the target protein. 
       FIGS. 5A to 5H  show the binding ability of antibodies screened by hybridoma to 293T cells. 
       FIGS. 6A to 6H  and  FIGS. 7A to 7F  show the binding ability of phage-screened antibodies to CHOS cells. 
     In the figures, 293T-QD012:293T transiently transfected with CLDN18.1. 
     293T-QD010:293T transiently transfected with CLDN18.2 
     Purpose: To verify the binding ability of antibody to claudin18.1 (non-specific binding) and claudin18.2 in 293-T cell line. 
     293T-QD210:293T-CLDN18.2-18.1 ECD1 transient cells (293T transiently transfected with CLDN18.2-18.1 ECD1, the first extracellular domain of CLDN 18.2 was replaced by the first extracellular domain of CLDN 18.1). 
     293T-QD211:293T-CLDN18.1-18.2 ECD1 transient cells ((293T transiently transfected with CLDN18.1-18.2 ECD1, the first extracellular domain of CLDN 18.1 was replaced by the first extracellular domain of CLDN 18.2). 
     Purpose: To verify that the antibody binding region is the domain encoded by claudin 18.2 exon-1 in 293-T cell line. 
     Hybridoma clone in 293T and phage clone in 293T similarly refer to the antibodies screened from hybridoma and phage based binding ability to different 293T transient cell lines, respectively. 
     The purpose of the experiment in 293T is the same as in CHO cell line, and the meanings of each curve and its representation are explained as follows. 
     1. The red curve (5) indicates the binding ability of the antibody to be tested to the 293T cell line transiently transfected with empty vector, that is, the negative control. 
     2. The blue curve (4) indicates the binding of the antibody to be tested to the 293T cell line transiently transfected with claudin 18.2 (293T-QD010) to detect the binding ability of the target protein. 
     3. The orange curve (3) indicates the binding of the antibody to be tested to the 293T cell line transiently transfected with claudin18.1 (293T-QD012) to detect the non-specific binding. 
     4. The dark green curve (2) indicates the binding of the antibody to be tested to the 293T cell line transiently transfected with claudin18.1-18.2 ECD1 (the first extracellular domain of CLDN18.1 was replaced by the first extracellular domain of 18.2, which was the region where the antibody binding site was designed) (293T-QD211), to verify the domain of the antibody binding site. 
     5. The light green curve (1) indicates the binding of the antibody to be tested to the 293T cell line transiently transfected with claudin 18.2-18.1 ECD1 (the first extracellular domain of CLDN18.2 was replaced by the first extracellular domain of 18.1, the former was the region where the antibody binding site was designed) (293T-QD210), to verify the necessity of this domain for antibody binding. 
       FIGS. 8A to 8H  show the binding ability of antibody to GA0006 tumor cells of gastric cancer PDX model. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Binding ratio of antibody to gastric cancer PDX model 
               
               
                 GA0006 tumor cells 
               
            
           
           
               
               
               
            
               
                   
                 Antibody number 
                 Ratio of antibody-bound cells 
               
               
                   
                   
               
               
                   
                 IMAB362 
                 85.56% 
               
               
                   
                 QP190191 
                 91.55% 
               
               
                   
                 QP192193 
                 92.37% 
               
               
                   
                 QP201202 
                 89.95% 
               
               
                   
                 QP207208 
                 91.66% 
               
               
                   
                 QP11091110 
                 92.37% 
               
               
                   
                 QP11131114 
                 90.31% 
               
               
                   
                 QP11151116 
                 91.66% 
               
               
                   
                   
               
            
           
         
       
     
     The above experimental results show that the antibodies provided by the present invention have better binding ability to proteins and lower non-specific binding. 
     In other embodiments, the present invention also provides a polynucleotide encoding the antibody as described above. The polynucleotides encoding the antibodies provided by the present invention, when provided in the form of DNA, may contain non-coding sequences that will be removed during subsequent transcription and editing, or may only contain sequences encoding sequences corresponding to the antibody provided by embodiments of the present invention and sequences necessary for protein expression. 
     The present invention also provides a pharmaceutical composition comprising the antibody described in any one of the above, and the pharmaceutical composition provided by the present invention may only contain any one or a combination of at least two of the antibodies provided in the embodiments. 
     It should be clear to those skilled in the art that, for pharmaceutical compositions, a pharmaceutically acceptable excipient is also included. Conventional excipients required to make powders or tablets and other dosage forms should be used as ingredients that should be added in the pharmaceutical process. 
     The present invention also provides a method for detecting whether CLDN exists in a biological sample, which comprises: 
     a step of administering an antibody according to any one of the above to a biological sample, wherein the antibody has a detectable label, and a step of detecting the presence or content of the detectable label. 
     Pharmacodynamic Test in Animal 
     a) Tumor tissues were collected from gastric cancer xenograft model GA0006 tumor-bearing mice, and cut into tumor masses with a diameter of 2-3 mm, which were inoculated subcutaneously at the right anterior scapula of Balb/c nude mice. 
     b) When the average tumor volume of Balb/c nude mice reached about 100 mm 3 , the mice were randomly divided into different groups (6 mice in each group). All animals were weighed, and the tumor volume was measured with vernier caliper. According to the tumor volume, random grouping method was adopted to ensure that the tumor volume among different groups was similar. The coefficient of variation (CV) of tumor volume in each group was calculated by the formula CV=SD/MTV×100%, which should be less than 40%. 
     Random grouping was done using StudyDirector™. The grouping day was DO, and the administration was started on the same day. The detailed administration method, dosage and route of administration are shown in the table below. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Administration parameters of animal pharmacodynamic test 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Number 
                   
                   
                   
                   
               
               
                   
                 of 
                 Administration 
                 Dose 
                 Mode of 
                 Administration 
               
               
                 Group 
                 animals 
                 group 
                 (mg/kg) 
                 administration 
                 cycle 
               
               
                   
               
               
                 1 
                 6 
                 Vehicle control 
                 — 
                 i.p. 
                 BIW × 4 weeks 
               
               
                 2 
                 6 
                 QP192193 
                 10 
                 i.p. 
                 BIW × 4 weeks 
               
               
                 3 
                 6 
                 QP207208 
                 10 
                 i.p. 
                 BIW × 4 weeks 
               
               
                 4 
                 6 
                 QP11151116 
                 10 
                 i.p. 
                 BIW × 4 weeks 
               
               
                 5 
                 6 
                 IMAB362 
                 10 
                 i.p. 
                 BIW × 4 weeks 
               
               
                   
               
            
           
         
       
     
     The dosage volume was 10 μL/g. 
     The meaning of antibody numbers in Tables 1 and 2, such as QP190191, denotes a combination of a heavy chain and a light chain. 
     c) After starting the administration, the body weight and tumor volume of mice were measured twice a week. The formula of calculation tumor volume: Tumor volume (mm 3 )=½×(a×b 2 ) (wherein, a denotes the long diameter and b denotes the short diameter). The experiment was terminated one week after the last administration, the mice were sacrificed, and the tumors were taken out, weighed, and photographed. The following analysis methods are selected for data analysis: 
     Relative tumor proliferation rate, T/C (%), that is, the percentage value of relative tumor volume or tumor weight between the treatment group and the control group at a certain time point. The calculation formula is: T/C %=TRTV/CRTV×100% (TRTV: average RTV of the treatment group; CRTV: average RTV of the control group; RTV=Vt/V0, V0 is the tumor volume of the animal at the time of grouping, and Vt is the tumor volume of the animals after treatment). 
     The relative tumor inhibition rate, TGI (%), is calculated as TGI %=(1-T/C)×100% (T and C are the relative tumor volume (RTV) of the treatment group and the control group at a certain time point, respectively). 
     The mean tumor volume of mice in PBS control group was 911.16±177.81 mm 3  on the 31st day after administration. The mean tumor volume of the antibodies QP192193 (10 mg/kg), QP207208 (10 mg/kg) and QP11151116 (10 mg/kg) was 580.97±67.97 mm 3 , 680.28±193.50 mm 3 , and 722.38±118.07 mm 3  on the 31st day after administration, respectively. The TGI of that was 36.34%, 25.34% and 20.72%, respectively. The mean tumor volume of control molecule IMAB362 (10 mg/kg) was 661.28±104.49 mm 3  on the 31st day after administration, and TGI was 27.42% (see the table below). All three screened molecules inhibited tumor growth to a certain extent, although there was no statistically significant difference. QP192193 showed a trend that it was better than the control molecule IMAB362, and QP207208 and QP11151116 also showed the same level of tumor inhibition ability as IMAB362. Moreover, the body weight of mice did not decrease significantly during the administration process, indicating that antibody molecules had no obvious toxic and side effects on mice. 
     
       
         
           
               
               
               
               
               
             
               
                   
               
               
                   
                 D0 Tumor 
                 D31 tumor 
                   
                 P value (t 
               
               
                 Groups 
                 Volume/mm 3   
                 volume/mm 3   
                 TGI % 
                 test) 
               
               
                   
               
             
            
               
                 PBS 
                 116.33 
                 911.16 
                 — 
                 — 
               
               
                 QP192193 
                 116.11 
                 580.97 
                 41.52% 
                 0.11 
               
               
                 QP207208 
                 116.06 
                 680.28 
                 29.01% 
                 0.40 
               
               
                 IMAB362 
                 116.34 
                 661.28 
                 31.44% 
                 0.25 
               
               
                 QP11151116 
                 116.22 
                 722.38 
                 23.74% 
                 0.40 
               
               
                   
               
            
           
         
       
     
     The analysis results of tumor weight are similar to those of tumor volume. The average tumor weight of mice in PBS control group was 722.57±176.32 mg on the 31st day after administration. The average tumor weights of the antibody molecules QP192193, QP207208, and QP11151116 treatment groups were 455.5±46.42 mg, 391.93±111.15 mg and 432.03±66.25 mg on the 31st day after the end of administration, respectively, and the TGI was 36.96%, 45.76% and 40.21%, respectively. The average tumor weight of control molecule IMAB362 treatment group was 435.78±91 mg on the 31st day after administration, and the TGI was 39.69%. (See the table below) 
     
       
         
           
               
               
               
               
               
             
               
                   
                   
               
               
                   
                   
                 D31 tumor 
                   
                 P value  
               
               
                   
                 Groups 
                 weight/g 
                 TGI/% 
                 (t test) 
               
               
                   
                   
               
             
            
               
                   
                 PBS 
                 0.722 
                 — 
                 — 
               
               
                   
                 QP192193 
                 0.455 
                 36.96% 
                 0.26 
               
               
                   
                 QP207208 
                 0.392 
                 45.76% 
                 0.12 
               
               
                   
                 IMAB362 
                 0.436 
                 39.69% 
                 0.21 
               
               
                   
                 QP11151116 
                 0.432 
                 40.21% 
                 0.20 
               
               
                   
                   
               
            
           
         
       
     
     Therefore, through the PDX animal pharmacodynamic test, we found that the selected antibody molecule antibody molecule had better or the same level of tumor suppressing ability in vivo than the control molecule IMAB362. 
     The experimental results are shown in  FIGS. 9 to 14 , in which the results of the pharmacodynamic test of antibody against gastric cancer PDX model GA0006 are shown in  FIG. 9 .  FIG. 10A  is the tumor growth curve of PBS group (negative control) after grouping.  FIG. 10B  is the tumor growth curve of QP192193 group after grouping.  FIG. 10C  is the tumor growth curve of QP207208 group after grouping.  FIG. 10D  is the tumor growth curve of QP11151116 group after grouping.  FIG. 10E  is the tumor growth curve of the control antibody IMAB362 group after grouping.  FIG. 11  shows the weight of tumor at D31 in each group of mice added with each antibody of the present invention and a control antibody.  FIG. 12  is a real shot diagram of the tumor volume of each experimental group.  FIG. 13  is the body weight curve of mice in each group.  FIG. 14  is the curve of body weight change rate of mice in each group. It can be seen from the experimental results that the invented antibody exhibits a better effect on inhibiting tumor growth than IMAB362 in animal pharmacodynamics in vivo.