Monoclonal antibody aiming at hepatitis B virus HBsAg as well as preparation and application thereof

Abstract

The invention discloses a monoclonal antibody aiming at hepatitis B virus HBsAg as well as preparation and application of the monoclonal antibody. The monoclonal antibody is composed of SY-1, SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, SY-11, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, SY-33, SY-34, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, SY-43, SY-44, SY-45, SY-46, SY-47, SY-48, SY-49 and SY-. The monoclonal antibody is composed of SY-1, SY-2, SY-3, SY-48, SY-49 and SY-. The monoclonal antibody obtained through screening is high in HBsAg binding capacity and has multiple biological functions and the capacity of recognizing different HBV genotypes.

Description

Technical Field

[0001] This application relates to the field of HBV antibody technology, specifically to monoclonal antibodies against hepatitis B virus HBsAg and their preparation and application. Background Technology

[0002] Hepatitis B virus (HBV) infection remains a major global public health problem. Chronic HBV infection can lead to a range of liver diseases, including chronic hepatitis B (CHB), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). Currently, the FDA-approved drugs for the treatment of chronic hepatitis B fall into two main categories: interferons and nucleoside analogs. These drugs exert their antiviral effects by modulating the host immune system and inhibiting viral replication, respectively. While existing drugs exhibit strong safety and antiviral activity, their therapeutic efficacy is quite limited. The HBsAg seroconversion rate with these drugs is less than 5%, failing to achieve clinical cure. Therefore, there is a need to develop more effective new drugs to improve the clinical management of HBV-related diseases.

[0003] Antibodies play a crucial role in preventing infection and helping the body clear viruses, and can be used as a natural, highly effective treatment for chronic hepatitis B. Antibodies targeting viral surface antigens can prevent viral entry into hepatocytes by blocking the interaction between the hepatitis B virus and its pro-receptor HSPG through the Fab region, or by being internalized into hepatocytes and subsequently blocking the release of the hepatitis B virus from infected cells to prevent a new wave of infection. On the other hand, antibodies can also interact with phagocytes through their Fc region, mediating the clearance of antigen-antibody immune complexes, i.e., the antibody ADCP effect. While traditional hepatitis B immunoglobulin (HBIG) is widely used clinically, it is mainly derived from the serum of hepatitis B vaccine recipients with high antibody titers, resulting in low production and significant differences in concentration and titer between different batches. Therefore, there is an urgent need to develop anti-hepatitis B virus monoclonal antibodies with higher titers and better specificity to provide new treatment options for chronic hepatitis B. Summary of the Invention

[0004] The purpose of this invention is to provide a monoclonal antibody against hepatitis B virus HBsAg, as well as its preparation and application.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: The first aspect of the present invention discloses a monoclonal antibody against hepatitis B virus HBsAg, wherein the monoclonal neutralizing antibody is SY-1, which is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence.

[0006] The second aspect of the present invention discloses a monoclonal antibody composition against hepatitis B virus HBsAg, characterized in that the monoclonal neutralizing antibody composition is SY-1 as described in claim 1 combined with monoclonal antibodies SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, SY-11, SY-12, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, and SY-2. 6. SY-27, SY-28, SY-29, SY-30, SY-31, SY-32, SY-33, SY-34, SY-35, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, S Any combination of Y-43, SY-44, SY-45, SY-46, SY-47, SY-48, SY-49, SY-50, SY-51, SY-52, SY-53, SY-54, SY-55, SY-56, SY-57; The SY-2 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, IQNSGST and GRLGHDFWSGYLRFDYNMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSDHS, DTS and LLSYSGVRV in sequence. The SY-3 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-5 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRF, AAS and QQYGNTPLCS in sequence. The SY-6 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLSDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNTPLCS in sequence. The SY-7 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, TSYDGSYT and AKEGNAFRYFGWLPDYQYYGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNSPLCS in sequence. The SY-8 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGTTT and AKEGNAFRYFGLLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRF, AGS and QQYGNSPLCS in sequence. The SY-9 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGTST and AKEGNAFRYFGFLPDYQYYGMDL in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRF, AAS and QQYGNSPLCS in sequence. The SY-10 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFTTYA, ISHDGNGK and AREFDCSGITCNTGRAFQY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSFNNEYH, WAS and QQYYSNPWT in sequence. The SY-11 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFQDTW, LSTQAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-12 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFQDTW, LSTQAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-13 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFRDVW, IKPKSGGGTS and ATFFQGYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQALQTPYT in sequence. The SY-14 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFRDVW, IKPKSGGGTS and ATFFQGAFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQSLQTPYT in sequence. The SY-15 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSNTW, IKSQPDGGTT and ATFFQGYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQSNGYNY, LGS and MQALQTPYT in sequence. The SY-16 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFTDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-17 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFTDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-18 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-19 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFDFTNAW, IRSNPNGGTA and VTQFLGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHRNGRSY, LGS and MQALQPPYS in sequence. The SY-20 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTNTYG, ISIGNSYK and AKLGVGYSSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are DSNIGNNY, KNN and AAWDNSLSGVI in sequence. The SY-21 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFILSDHY, SRNKANSYTT and VRSILVTTGGNYFEC in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QGISSS, DAS and QQSKSYPLT in sequence. The SY-22 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYA, IDTSTGNP and VRGSAVHILRFFDWRRFDD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGSNF, DVN and CSYAASYTWV in sequence. The SY-26 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence. The SY-27 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence. The SY-28 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYNGLRV in sequence. The SY-29 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYY, VQNSGNT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-30 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYY, VQNSGNI and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVSNGHS, DTS and LLSYSGVRV in sequence. The SY-31 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, IQNSGNT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-32 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYF, IQNSGST and GRQGHDFWSGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTN and LLSYSGVRV in sequence. The SY-33 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDI, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV, respectively. The SY-34 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-35 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, IQNSGST and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-36 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-37 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, MQNTGAT and GRMGHDFWSGYLRFDYNSDI, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGGVTSGHS, DTS and LLSYSGVRV, respectively. The SY-38 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTRGYA, ISGSGLST and AKDPVSPVTVVGTFFQH in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGYNY, EVT and SSYAGSNTLGV in sequence. The SY-39 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLNT and AKDPVTPITVVGTFFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYNY, EVT and SSFAGSSNLGV in sequence. The SY-40 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLST and AKDPVTPITVVGTYFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYNY, EVT and SSFAGSSNLGV in sequence. The SY-41 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLNT and AKDPVTPITVVGTFFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYSY, EVT and SSFAGSSNLGV in sequence. The SY-43 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are DFSFSSHA, ISYDGGTT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRY, AAS and QQYGNSPLCS in sequence. The SY-44 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYYGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QNINSRY, AAS and QQYGNSPLCS in sequence. The SY-45 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNTPLCS in sequence. The SY-46 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFKDTW, LSTEAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-47 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFDFNHVW, IKSKSDGATT and ATFFHAYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are RSLLQRNGDNY, LGT and QGLQTPYT in sequence. The SY-48 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDVW, IKPKSGGGTS and ATFFQGYFDY, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQALQTPYT, respectively. The SY-49 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDAW, IKTTALGATS and TTLFGGAFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTVMNRNGYNY, LSS and MQSVQTPYT in sequence. The SY-50 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are EFTLSRHL, ISIGSSYI and VRLGVGFTSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are NSNIGSNY, KNN and AAWDNSLSGVA in sequence. The SY-51 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFNRYG, ISIGSSYK and AKLGVGYSSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are DSNIGNNY, KNN and AAWDNSLGGVV in sequence. The SY-52 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFSDHY, TRNRANSYRT and VRGPLVGKTGFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QGIRNY, TVS and LQHSSYPYS in sequence. The SY-53 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are HFTFSSYA, IWYDGTYK and AKSLTGALEG in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLNSDDGNTY, TVS and MHRVEFPRT in sequence. The SY-54 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYSFTNYA, IDTYTGNP and ARGSAVHILQFFDWRRFDD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGYNF, DVS and CSYAGSYTWV in sequence. The SY-55 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GDTFSSYT, INGFLGIA and ARGDAGAVRLLYYNYAMDL, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQSDGKTY, EVS and MQTIQLPLT, respectively. The SY-56 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GGTFRSYG, IVPIFGIS and VRGGAVAGAGSAGEGIFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSVSKN, GAS and QQHHSWPPFT in sequence. The SY-57 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTKFG, ITPYNGNR and SRDGDRAAVSSF, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are ESLFFDSLNKNY, WAS and QQYIMSPWT, respectively.

[0007] It should be noted that the key to this invention lies in isolating and expanding plasmablasts from peripheral blood mononuclear cells (PBMCs) collected on the 7th day after vaccination, ultimately obtaining 52 fully human monoclonal neutralizing antibodies, namely SY-1, SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, SY-11, SY-12, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, SY-26, SY-27, and SY- 28. SY-29, SY-30, SY-31, SY-32, SY-33, SY-34, SY-35, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, SY-43, SY-44, SY-45, SY-46, SY-47, SY-48, SY-49, SY-50, SY-51, SY-52, SY-53, SY-54, SY-55, SY-56, SY-57, 52 strains of monoclonal neutralizing antibodies have binding activity and antiviral function to HBsAg proteins of different HBV genotypes.

[0008] In one implementation of the present invention, the heavy chain variable region sequence of SY-1 is Seq ID No.1, and the light chain variable region sequence is Seq ID No.2; The heavy chain variable region sequence of SY-2 is Seq ID No. 3, and the light chain variable region sequence is Seq ID No. 4; The heavy chain variable region sequence of SY-3 is Seq ID No. 5, and the light chain variable region sequence is Seq ID No. 6; The heavy chain variable region sequence of SY-4 is Seq ID No. 7, and the light chain variable region sequence is Seq ID No. 8; The heavy chain variable region sequence of SY-5 is Seq ID No. 9, and the light chain variable region sequence is Seq ID No. 10; The heavy chain variable region sequence of SY-6 is Seq ID No. 11, and the light chain variable region sequence is Seq ID No. 12; The heavy chain variable region sequence of SY-7 is Seq ID No. 13, and the light chain variable region sequence is Seq ID No. 14; The heavy chain variable region sequence of SY-8 is Seq ID No. 15, and the light chain variable region sequence is Seq ID No. 16; The heavy chain variable region sequence of SY-9 is Seq ID No. 17, and the light chain variable region sequence is Seq ID No. 18; The heavy chain variable region sequence of SY-10 is Seq ID No. 19, and the light chain variable region sequence is Seq ID No. 20; The heavy chain variable region sequence of SY-11 is Seq ID No. 21, and the light chain variable region sequence is Seq ID No. 22; The heavy chain variable region sequence of SY-12 is Seq ID No. 23, and the light chain variable region sequence is Seq ID No. 24; The heavy chain variable region sequence of SY-13 is Seq ID No. 25, and the light chain variable region sequence is Seq ID No. 26; The heavy chain variable region sequence of SY-14 is Seq ID No. 27, and the light chain variable region sequence is Seq ID No. 28; The heavy chain variable region sequence of SY-15 is Seq ID No. 29, and the light chain variable region sequence is Seq ID No. 30; The heavy chain variable region sequence of SY-16 is Seq ID No. 31, and the light chain variable region sequence is Seq ID No. 32; The heavy chain variable region sequence of SY-17 is Seq ID No. 33, and the light chain variable region sequence is Seq ID No. 34; The heavy chain variable region sequence of SY-18 is Seq ID No. 35, and the light chain variable region sequence is Seq ID No. 36; The heavy chain variable region sequence of SY-19 is Seq ID No. 37, and the light chain variable region sequence is Seq ID No. 38; The heavy chain variable region sequence of SY-20 is Seq ID No. 39, and the light chain variable region sequence is Seq ID No. 40; The heavy chain variable region sequence of SY-21 is Seq ID No. 41, and the light chain variable region sequence is Seq ID No. 42; The heavy chain variable region sequence of SY-22 is Seq ID No. 43, and the light chain variable region sequence is Seq ID No. 44; The heavy chain variable region sequence of SY-26 is Seq ID No. 45, and the light chain variable region sequence is Seq ID No. 46; The heavy chain variable region sequence of SY-27 is Seq ID No. 47, and the light chain variable region sequence is Seq ID No. 48; The heavy chain variable region sequence of SY-28 is Seq ID No. 49, and the light chain variable region sequence is Seq ID No. 50; The heavy chain variable region sequence of SY-29 is Seq ID No. 51, and the light chain variable region sequence is Seq ID No. 52; The heavy chain variable region sequence of SY-30 is Seq ID No. 53, and the light chain variable region sequence is Seq ID No. 54; The heavy chain variable region sequence of SY-31 is Seq ID No. 55, and the light chain variable region sequence is Seq ID No. 56; The heavy chain variable region sequence of SY-32 is Seq ID No. 57, and the light chain variable region sequence is Seq ID No. 58; The heavy chain variable region sequence of SY-33 is Seq ID No. 59, and the light chain variable region sequence is Seq ID No. 60; The heavy chain variable region sequence of SY-34 is Seq ID No. 61, and the light chain variable region sequence is Seq ID No. 62; The heavy chain variable region sequence of SY-35 is Seq ID No. 63, and the light chain variable region sequence is Seq ID No. 64; The heavy chain variable region sequence of SY-36 is Seq ID No. 65, and the light chain variable region sequence is Seq ID No. 66; The heavy chain variable region sequence of SY-37 is Seq ID No. 67, and the light chain variable region sequence is Seq ID No. 68; The heavy chain variable region sequence of SY-38 is Seq ID No. 69, and the light chain variable region sequence is Seq ID No. 70; The heavy chain variable region sequence of SY-39 is Seq ID No. 71, and the light chain variable region sequence is Seq ID No. 72; The heavy chain variable region sequence of SY-40 is Seq ID No. 73, and the light chain variable region sequence is Seq ID No. 74; The heavy chain variable region sequence of SY-41 is Seq ID No. 75, and the light chain variable region sequence is Seq ID No. 76; The heavy chain variable region sequence of SY-43 is Seq ID No. 77, and the light chain variable region sequence is Seq ID No. 78; The heavy chain variable region sequence of SY-44 is Seq ID No. 79, and the light chain variable region sequence is Seq ID No. 80; The heavy chain variable region sequence of SY-45 is Seq ID No. 81, and the light chain variable region sequence is Seq ID No. 82; The heavy chain variable region sequence of SY-46 is Seq ID No. 83, and the light chain variable region sequence is Seq ID No. 84; The heavy chain variable region sequence of SY-47 is Seq ID No. 85, and the light chain variable region sequence is Seq ID No. 86; The heavy chain variable region sequence of SY-48 is Seq ID No. 87, and the light chain variable region sequence is Seq ID No. 88; The heavy chain variable region sequence of SY-49 is Seq ID No. 89, and the light chain variable region sequence is Seq ID No. 90; The heavy chain variable region sequence of SY-50 is Seq ID No. 91, and the light chain variable region sequence is Seq ID No. 92; The heavy chain variable region sequence of SY-51 is Seq ID No. 93, and the light chain variable region sequence is Seq ID No. 94; The heavy chain variable region sequence of SY-52 is Seq ID No. 95, and the light chain variable region sequence is Seq ID No. 96; The heavy chain variable region sequence of SY-53 is Seq ID No. 97, and the light chain variable region sequence is Seq ID No. 98; The heavy chain variable region sequence of SY-54 is Seq ID No. 99, and the light chain variable region sequence is Seq ID No. 100; The heavy chain variable region sequence of SY-55 is Seq ID No. 101, and the light chain variable region sequence is Seq ID No. 102; The heavy chain variable region sequence of SY-56 is Seq ID No. 103, and the light chain variable region sequence is Seq ID No. 104; The heavy chain variable region sequence of SY-57 is Seq ID No. 105, and the light chain variable region sequence is the sequence shown in Seq ID No. 106.

[0009] It should be noted that the heavy chain variable regions and light chain variable regions of the above specific sequences are only the specific monoclonal neutralizing antibody sequences used in one implementation of the present invention. It can be understood that for monoclonal neutralizing antibodies, the regions that affect their precise complementarity with the antigenic determinant are complementarity-determining regions (CDRs), specifically CDR1, CDR2, and CDR3 of the heavy chain, and CDR1, CDR2, and CDR3 of the light chain. Therefore, as long as the heavy chain and light chain CDR1, CDR2, and CDR3 sequences of the present invention remain unchanged, the function and role of the hepatitis B virus monoclonal neutralizing antibody of the present invention can be basically realized. In other words, the specific sequence of the hepatitis B virus monoclonal neutralizing antibody of the present invention is not limited to the heavy chain variable regions and light chain variable regions of the above specific sequences.

[0010] A third aspect of the present invention discloses a biomaterial selected from one of (A)-(B) below: (A) A nucleic acid molecule encoding the monoclonal antibody against hepatitis B virus HBsAg, or the monoclonal antibody composition against hepatitis B virus HBsAg; (B) Recombinant plasmids and recombinant cells containing the nucleic acid molecules described in (A).

[0011] In one implementation of the present invention, the nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 1 is shown in Seq ID No. 107, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 2 is shown in Seq ID No. 108. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 3 is shown in Seq ID No. 109, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 4 is shown in Seq ID No. 110. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 5 is shown in Seq ID No. 111, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 6 is shown in Seq ID No. 112. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 7 is shown in Seq ID No. 113, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 8 is shown in Seq ID No. 114. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 9 is shown in Seq ID No. 115, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 10 is shown in Seq ID No. 116. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 11 is shown in Seq ID No. 117, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 12 is shown in Seq ID No. 118. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 13 is shown in Seq ID No. 119, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 14 is shown in Seq ID No. 120. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 15 is shown in Seq ID No. 121, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 16 is shown in Seq ID No. 122. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 17 is shown in Seq ID No. 123, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 18 is shown in Seq ID No. 124. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 19 is shown in Seq ID No. 125, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 20 is shown in Seq ID No. 126. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 21 is shown in Seq ID No. 127, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 22 is shown in Seq ID No. 128. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 23 is shown in Seq ID No. 129, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 24 is shown in Seq ID No. 130. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 25 is shown in Seq ID No. 131, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 26 is shown in Seq ID No. 132. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 27 is shown in Seq ID No. 133, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 28 is shown in Seq ID No. 134. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 29 is shown in Seq ID No. 135, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 30 is shown in Seq ID No. 136. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 31 is shown in Seq ID No. 137, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 32 is shown in Seq ID No. 138. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 33 is shown in Seq ID No. 139, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 34 is shown in Seq ID No. 140. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 35 is shown in Seq ID No. 141, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 36 is shown in Seq ID No. 142. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 37 is shown in Seq ID No. 143, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 38 is shown in Seq ID No. 144. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 39 is shown in Seq ID No. 145, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 40 is shown in Seq ID No. 146. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 41 is shown in Seq ID No. 147, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 42 is shown in Seq ID No. 148. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 43 is shown in Seq ID No. 149, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 44 is shown in Seq ID No. 150. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 45 is shown in Seq ID No. 151, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 46 is shown in Seq ID No. 152. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 47 is shown in Seq ID No. 153, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 48 is shown in Seq ID No. 154. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 49 is shown in Seq ID No. 155, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 50 is shown in Seq ID No. 156. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 51 is shown in Seq ID No. 157, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 52 is shown in Seq ID No. 158. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 53 is shown in Seq ID No. 159, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 54 is shown in Seq ID No. 160. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 55 is shown in Seq ID No. 161, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 56 is shown in Seq ID No. 162. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 57 is shown in Seq ID No. 163, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 58 is shown in Seq ID No. 164. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 59 is shown in Seq ID No. 165, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 60 is shown in Seq ID No. 166. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 61 is shown in Seq ID No. 167, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 62 is shown in Seq ID No. 168. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 63 is shown in Seq ID No. 169, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 64 is shown in Seq ID No. 170. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 65 is shown in Seq ID No. 171, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 66 is shown in Seq ID No. 172. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 67 is shown in Seq ID No. 173, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 68 is shown in Seq ID No. 174. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 69 is shown in Seq ID No. 175, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 70 is shown in Seq ID No. 176. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 71 is shown in Seq ID No. 177, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 72 is shown in Seq ID No. 178. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 73 is shown in Seq ID No. 179, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 74 is shown in Seq ID No. 180. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 75 is shown in Seq ID No. 181, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 76 is shown in Seq ID No. 182. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 77 is shown in Seq ID No. 183, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 78 is shown in Seq ID No. 184. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 79 is shown in Seq ID No. 185, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 80 is shown in Seq ID No. 186. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 81 is shown in Seq ID No. 187, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 82 is shown in Seq ID No. 188. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 83 is shown in Seq ID No. 189, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 84 is shown in Seq ID No. 190. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 85 is shown in Seq ID No. 191, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 86 is shown in Seq ID No. 192. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 87 is shown in Seq ID No. 193, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 88 is shown in Seq ID No. 194. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 89 is shown in Seq ID No. 195, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 90 is shown in Seq ID No. 196. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 91 is shown in Seq ID No. 197, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 92 is shown in Seq ID No. 198. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 93 is shown in Seq ID No. 199, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 94 is shown in Seq ID No. 200. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 95 is shown in Seq ID No. 201, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 96 is shown in Seq ID No. 202. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 97 is shown in Seq ID No. 203, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 98 is shown in Seq ID No. 204. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 99 is shown in Seq ID No. 205, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 100 is shown in Seq ID No. 206. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 101 is shown in Seq ID No. 207, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 102 is shown in Seq ID No. 208. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 103 is shown in Seq ID No. 209, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 104 is shown in Seq ID No. 210. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 105 is shown in Seq ID No. 211, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 106 is shown in Seq ID No. 212.

[0012] It should be noted that the specific nucleic acid sequences mentioned above are only the nucleic acid sequences used in one implementation of the present invention. It can be understood that there can be multiple codons for one amino acid. Therefore, based on the degeneracy of codons, in addition to the nucleic acid sequences defined above, there can be several other nucleic acid sequences that encode the same heavy or light chain, all of which are within the scope of protection of the present invention, while ensuring that the coding sequence remains unchanged.

[0013] The fourth aspect of the present invention discloses a method for preparing the monoclonal antibody against hepatitis B virus HBsAg, comprising: expressing the protein of the recombinant plasmid using the recombinant cells, extracting and purifying the expressed protein, thereby obtaining the monoclonal antibody against hepatitis B virus HBsAg.

[0014] It should be noted that the recombinant plasmids of the present invention are designed to effectively express the nucleic acid fragments of the present invention, thereby obtaining the corresponding heavy chain, light chain, or hepatitis B virus monoclonal neutralizing antibodies; therefore, in principle, any vector capable of transfecting the nucleic acid fragments into host cells for nucleic acid expression can be used in the present invention.

[0015] It should be noted that the recombinant cells of the present invention refer to host cells transfected with the nucleic acid fragments or recombinant plasmids of the present invention; generally, the heavy chain, light chain, or hepatitis B virus monoclonal neutralizing antibodies of the present invention can be obtained by directly culturing such host cells.

[0016] The fifth aspect of the present invention discloses the hepatitis B virus monoclonal neutralizing antibody, or the monoclonal antibody composition against hepatitis B virus HBsAg, or the use of the biological material in the preparation of hepatitis B virus prevention and treatment drugs or hepatitis B virus detection reagents.

[0017] It should be noted that the hepatitis B virus monoclonal neutralizing antibody of the present invention has cross-binding activity and antiviral function with HBsAg proteins of different HBV genotypes; therefore, it can be used to prepare corresponding drugs for the prevention and treatment of hepatitis B virus. As for nucleic acid fragments, recombinant plasmids, and recombinant cells, these can be used as raw materials for the preparation of hepatitis B virus monoclonal neutralizing antibodies, thereby being used to prepare hepatitis B virus prevention and treatment drugs or hepatitis B virus detection reagents.

[0018] The sixth aspect of the present invention discloses a hepatitis B virus prevention and treatment drug containing the aforementioned monoclonal antibody against hepatitis B virus HBsAg, or the aforementioned monoclonal antibody composition against hepatitis B virus HBsAg.

[0019] The seventh aspect of the present invention discloses a hepatitis B virus detection reagent containing the aforementioned monoclonal antibody against hepatitis B virus HBsAg, or the aforementioned monoclonal antibody composition against hepatitis B virus HBsAg.

[0020] Due to the adoption of the above technical solutions, the beneficial effects of the present invention are as follows: The 52 monoclonal neutralizing antibodies against hepatitis B virus of the present invention are HBsAg-specific fully human antibodies, and some of the antibodies have broad-spectrum neutralizing activity and ADCP activity, providing a new option for the prevention and treatment of hepatitis B virus.

[0021] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only, and are not intended to limit this disclosure. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 The results of the binding experiment between 52 monoclonal neutralizing antibodies and genotype A HBsAg protein in this embodiment of the invention are shown.

[0024] Figure 2 The results of the binding experiments of 52 monoclonal neutralizing antibodies to genotype C HBsAg protein in this embodiment of the invention are shown.

[0025] Figure 3 The results of the neutralization experiment of 52 monoclonal neutralizing antibodies in the embodiments of the present invention are shown.

[0026] Figure 4 The ADCP experimental results of 52 monoclonal neutralizing antibodies in this embodiment of the invention are shown.

[0027] Figure 5 The results of flow cytometry experiments in this embodiment of the invention show the binding of 5 monoclonal neutralizing antibodies to 10 different HBV genotypes HBsAg. Detailed Implementation

[0028] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Experimental methods in the following embodiments that do not specify specific conditions are generally determined according to national standards. Experimental materials in the following embodiments that do not specify the source are all commercially available raw materials. The equipment used in each step of the following embodiments are all conventional equipment. If there is no corresponding national standard, it is carried out according to general international standards, conventional conditions, or conditions recommended by the manufacturer. Unless otherwise stated, all parts are parts by weight, and all percentages are percentages by mass. Unless otherwise defined or stated, all professional and scientific terms used in the present invention have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the methods of the present invention.

[0029] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but this is not intended to limit the scope of the invention. Example 1: Antibody screening, expression, and purification

[0030] This study was approved by the Ethics Committee of the Third People's Hospital of Shenzhen (Approval No.: 2021-030). Participants provided written informed consent for sample collection and subsequent analysis.

[0031] 1. This example specifically collected one sample, and the sample collection method is as follows: Peripheral blood mononuclear cell isolation: Collect 10 mL of venous blood using an anticoagulant tube, transfer it to a 50 mL centrifuge tube, dilute with 10 mL of PBS solution, and mix gently. Take two 15 mL centrifuge tubes and add 5 mL of Ficoll separation solution to each. Then add 10 mL of diluted blood to the supernatant of the Ficoll separation solution. Centrifuge at 2000 rpm for 20 minutes. Transfer the leukocyte layer to a clean 15 mL centrifuge tube. Add PBS to 10 mL, centrifuge at 1500 rpm for 10 minutes, discard the supernatant, and collect the cells for subsequent experiments.

[0032] 2. Isolation of monoclonal antibodies from peripheral blood clone-expanded plasmablasts Fresh peripheral blood mononuclear cells were isolated and washed twice with 10 mL of PBS. A mixture of CD19-PE-Cy7, CD20-APC-H7, CD3-APC-H7, CD27-PE, and CD38-APC (all from BD Biosciences) was added to 100 μL of staining buffer (PBS + 2% fetal bovine serum) to resuspend the peripheral blood mononuclear cells, and staining was performed at 4°C for 30 minutes. After washing twice with PBS, single activated plasmablasts of CD19+CD20-CD3-CD27^hiCD38^hi were sorted using a BDAria II sorting flow cytometer.

[0033] Single activated plasmablasts were sorted into 96-well PCR plates containing lysis buffer, and then RT-PCR and nested PCR were performed according to the method described in the literature (LiaoHX, Levesque MC, Nagel A, Dixon A, Zhang R, Walter E, et al. High-throughputisolation of immunoglobulin genes from single human B cells and expression asmonoclonal antibodies. Journal of virological methods. 2009;158:171-9.) to amplify the variable regions of the heavy and light chains, respectively. The PCR amplification products were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing, and the antibodies obtained were named SY-1, SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, S Y-11, SY-12, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, SY-26, SY-27, SY- 28. SY-29, SY-30, SY-31, SY-32, SY-33, SY-34, SY-35, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, SY-4 3. SY-44, SY-45, SY-46, SY-47, SY-48, SY-49, SY-50, SY-51, SY-52, SY-53, SY-54, SY-55, SY-56, SY-57.

[0034] The antibody variable region sequence obtained from sequencing was sent to GenScript Biotech for synthesis. GenScript then cloned the variable regions of the antibody heavy and light chains into the full-length IgG1 heavy and light chain expression vectors pCDNA3.4 (GenScript Biotech), respectively, and prepared large quantities of antibody heavy and light chain plasmids. After obtaining the plasmids prepared by GenScript, the paired heavy and light chain expression plasmids were co-transfected into 293F cells (500 mL as an example) using PEI transfection reagent for expression. The monoclonal antibody was then purified from the culture supernatant using a protein A adsorption column. The specific steps are as follows: 293F cells were cultured in an 8% CO2, 37℃ incubator. The cell concentration was adjusted to 1.2 × 10⁶ cells / mL, and cultured for another 2 hours. Solution A was prepared by adding 250 μg of antibody heavy chain plasmid and 250 μg of antibody light chain plasmid to 12.5 mL of Opti-MEM (31985070, Gibco). Solution B was prepared by adding 2.5 mL of 1 mg / mL PEI transfection reagent (24885-2, Polysciences) to 12.5 mL of Opti-MEM and incubating for 5 minutes. Solutions A and B were mixed and incubated for 20 minutes to obtain the AB mixture. 25 mL of the AB mixture was added dropwise to 500 mL of 293F cells, shaking constantly while adding. The cells were cultured for another 5 days. The 293F cells were then centrifuged at 3000g for 20 minutes, and the supernatant was collected and filtered through a 0.45 μm filter. Protein was then opened. The Protein A gravity column was capped, and gravity was used to allow the 20% ethanol solution in the column to flow out completely. The Protein A gravity column was equilibrated with 5 column volumes of 10 mM PBS solution. Filtered cell supernatant was added to the Protein A gravity column and allowed to flow out under gravity. The Protein A gravity column was washed with 3 column volumes of PBS solution, and then eluted with 5 volume volumes of 0.1 M glycine-hydrochloric acid solution (pH=3.0). The eluent was placed in a 30 kDa ultrafiltration concentration tube, filled with PBS, and centrifuged at 3500 rpm for 40 minutes at 4°C. The waste liquid in the collection tube was discarded, 20 mL of PBS solution was added, and the tube was centrifuged at 3500 rpm for 40 minutes at 4°C. The concentrated and replaced antibody solution was then collected, and the antibody protein concentration was measured. Example 2: Enzyme-linked immunosorbent assay (ELISA) to verify the binding ability of 52 antibodies.

[0035] In Example 1, single activated plasmablasts were isolated from the peripheral blood of vaccine recipients. Fifty-two fully human monoclonal antibodies were ultimately obtained using RT-PCR and nested PCR techniques, namely SY-1, SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, SY-11, SY-12, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, SY-26, SY-27, and SY-28. , SY-29, SY-30, SY-31, SY-32, SY-33, SY-34, SY-35, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, SY-43 , SY-44, SY-45, SY-46, SY-47, SY-48, SY-49, SY-50, SY-51, SY-52, SY-53, SY-54, SY-55, SY-56, SY-57.

[0036] The heavy chain and light chain variable region sequences of the 52 monoclonal neutralizing antibodies isolated in this example, and the sequencing results are shown in the sequence listing.

[0037] The binding affinity of 52 HBsAg monoclonal antibodies obtained to hepatitis B virus (HBsAg) was detected using an ELISA assay. HBsAg proteins of genotype A and genotype C (Shenzhen Kangtai Biological, Shenzhen Immuth Company) were coated in 96-well ELISA plates at 2 μg / mL and incubated overnight at 4°C, 100 μL per well. Wash 5 times with PBST (PBS solution containing 0.5% Tween-20); block with blocking buffer at room temperature for 1 hour, 150 μL per well, then wash 5 times with PBST. Blocking buffer formulation: 5% skim milk + 2% BSA (prepared with PBS). All subsequent antibody dilution buffer formulations are the same as the blocking buffer. Dilute the antibody to be tested to 10 μg / mL with blocking buffer, 100 μL per well, incubate at 37°C for 1 hour, then wash 5 times with PBST. Add HRP-goat anti-human IgG (ZB-2304, Zhongshan Jinqiao) at a ratio of 1:5000, incubate at 37°C for 1 hour, then wash 5 times with PBST. Mix chromogenic solution A and solution B (Sangon Biotech) at a ratio of 1:1, 100 μL per well, incubate at room temperature in the dark for 20 minutes; then stop the reaction with 50 μL of 2M H2SO4. Measure the optical density at 450 nm (OD) using a Varioskan LUX multimodal microplate reader (Thermo Scientific).

[0038] The binding activity of 52 monoclonal antibodies to the HBV surface antigen protein HBsAg was analyzed by ELISA. The results are as follows: Figure 1 and Figure 2 As shown, all 52 monoclonal antibodies could bind to HBsAg of HBV genotype A, and their binding ability was relatively strong. Among them, 50 monoclonal antibodies could cross-bind to HBsAg of HBV genotype C. Example 3: Neutralization capacity assay of 52 antibody strains

[0039] HepG2-AD38 cells and HepG2-NTCP cells were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin and streptomycin at 37°C in a 5% CO2 incubator.

[0040] Preparation of live HBV virus: When the confluence of HepG2-AD38 cells in the T75 cell culture flask reaches approximately 80%, the culture medium is discarded, and fresh culture medium is added. HBV virus particles are collected every 2 days for a total of 5 collections. The virus-containing cell culture medium is transferred to a 50 mL centrifuge tube, centrifuged at 3500 rpm for 20 minutes, and the supernatant is filtered through a 0.45 μm filter. The collected virus is then ultra-highly purified with 15% sucrose solution and centrifuged at 130,000 g for 16 hours. The supernatant is discarded, and the virus is resuspended in serum-free DMEM medium, mixed well, aliquoted, and stored at -80°C for later use.

[0041] To determine antibody neutralizing activity, serially diluted monoclonal antibodies were incubated with HBV dilution at 37°C for 1 hour. The mixture was then added to 96-well plates containing HepG2-NTCP cells and incubated at 37°C with 5% CO2 for 24 hours, after which the culture medium was replaced. On day 7 post-infection, the supernatant was collected, and HBeAg levels were detected using ELISA (KHB diagnostic kit). The HBeAg secretion inhibition rate was calculated using GraphPad Prism 8.0 software, employing a logarithmic (inhibitor) and standardized response-variable slope (four-parameter) model to calculate the 50% inhibitory concentration (IC50).

[0042] The neutralizing activity of 52 antibody strains against live HBV genotype D was detected by an HBV live virus neutralization assay. The results are as follows: Figure 3 As shown, 43 antibodies have a neutralizing effect on live HBV of genotype D and are HBV neutralizing antibodies. Example 4: Detection of ADCP activity of 52 antibody strains

[0043] THP-1 cells were digested and dispersed into single cells. After counting, the cell concentration was adjusted and seeded into 24-well plates with 2 x 10⁵ cells per well, 500 μL, and cultured at 37°C for 24 hours in a 5% CO₂ incubator. The antibody to be tested was diluted to a concentration of 20 μg / mL with RPMI 1640 medium containing 10% FBS. The diluted antibody was mixed with an equal volume of HBsAg protein (1 μg / mL) and incubated at 37°C for 1 hour. Aspirate 250 μL of cell culture medium and add 250 μL of the incubated antibody-antigen mixture to THP-cells. Incubate at 37°C for 2 hours in a 5% CO2 incubator. Discard the supernatant, wash the cells once with sterile PBS, add 200 μL of cell lysis buffer (Western and IP cell lysis buffer, Beyotime) to each well, vortex for 1 minute, and lyse at 4°C for 15 minutes. Add 200 μL of PBS and measure the lysis buffer diluted to 100 μL using an HBsAg ELISA kit (Beijing Wantai Biotech). Measure the optical density at 450 nm (OD) using a Varioskan LUX multimodal microplate reader (Thermo Scientific).

[0044] Besides neutralization, Fc-mediated non-neutralizing functions also contribute to antibody clearance of HBV, such as antibody-dependent phagocytosis (ADCP). Using THP-1 cells as effector cells, the biological activity of the antibody was tested in the presence of HBsAg particles. Results are as follows... Figure 4 As shown, 46 antibodies possess ADCP activity. Example 5: Detection of the broad-spectrum recognition ability of five monoclonal antibodies using flow cytometry

[0045] This embodiment uses flow cytometry to detect the intracellular expression of HBsAg in cells that recognize different HBV genotypes using 52 monoclonal antibodies, as detailed below: First, HBsAg protein expression plasmids for different HBV genotypes A, B, C, D, E, F, G, H, I, and J were transfected into 293T cells, with cells transfected with the empty Vector vector serving as a control. After 48 hours, cells were digested with 0.5% trypsin and the process was terminated by adding complete DMEM medium containing 10% FBS. Cells were gently pipetted and observed under a microscope until more than 90% of the cells were in a single-cell state. The single-cell suspension was transferred to a 15mL centrifuge tube and centrifuged at 400g for 5 minutes. The supernatant was discarded, and 3mL of complete DMEM medium was added. 20μL of the cell suspension was taken, and 20μL of AO / PI dye (RE010212, Countstar) was added. The viable cell count was determined at 5×10⁵ cells / test. Cells were fixed and permeabilized using a cell fixation / permeabilization kit (554714, BD Biosciences), centrifuged at 400g for 5 minutes, and the supernatant was discarded. Then, 5 μg / mL of the test antibody was added to each flow cytometry tube, and incubated at 4°C for 30 minutes. After washing, the cells were stained with goat anti-human secondary antibody (A11013, Life Technologies) conjugated to Alexa Fluor 647, and incubated at 4°C for 30 minutes. After washing, the cells were collected using a FACSymphony™ A3 flow cytometer (BD Biososciences), and the data were analyzed using FlowJo software V10.9 (BD Biososciences). The experimental results are as follows: Figure 5 As shown.

[0046] The results showed that the antibodies SY-34, SY-40, SY-46, SY-48, and SY-54 could clearly detect intracellularly expressed HBsAg and could widely identify HBsAg from different HBV genotypes A, B, C, D, E, F, G, H, I, and J.

[0047] In summary, the 52 monoclonal antibodies screened in this invention have a strong binding ability to the surface antigen protein HBsAg of hepatitis B virus. Some antibodies have neutralizing and ADCP activities, and can cross-recognize HBsAg of 10 types of hepatitis B virus, exhibiting good broad-spectrum and antiviral activity, providing a new option for the prevention and treatment of hepatitis B virus.

[0048] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A monoclonal antibody against hepatitis B virus HBsAg, characterized in that, The monoclonal neutralizing antibody is SY-1, which consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence.

2. A monoclonal antibody composition targeting hepatitis B virus HBsAg, characterized in that, The monoclonal neutralizing antibody composition is SY-1 according to claim 1 and monoclonal antibodies SY-2, SY-3, SY-5, SY-6, SY-7, SY-8, SY-9, SY-10, SY-11, SY-12, SY-13, SY-14, SY-15, SY-16, SY-17, SY-18, SY-19, SY-20, SY-21, SY-22, SY-26, SY-27, SY-28, SY-29, SY-30, SY-31, SY-32, SY-33, SY-34, SY-35, SY-36, SY-37, SY-38, SY-39, SY-40, SY-41, SY-43, SY-44 , any combination of SY-45, SY-46, SY-47, SY-48, SY-49, SY-50, SY-51, SY-52, SY-53, SY-54, SY-55, SY-56, and SY-57; The SY-2 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, IQNSGST and GRLGHDFWSGYLRFDYNMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSDHS, DTS and LLSYSGVRV in sequence. The SY-3 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-5 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRF, AAS and QQYGNTPLCS in sequence. The SY-6 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLSDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNTPLCS in sequence. The SY-7 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, TSYDGSYT and AKEGNAFRYFGWLPDYQYYGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNSPLCS in sequence. The SY-8 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGTTT and AKEGNAFRYFGLLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRF, AGS and QQYGNSPLCS in sequence. The SY-9 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGTST and AKEGNAFRYFGFLPDYQYYGMDL in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRF, AAS and QQYGNSPLCS in sequence. The SY-10 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFTTYA, ISHDGNGK and AREFDCSGITCNTGRAFQY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSFNNEYH, WAS and QQYYSNPWT in sequence. The SY-11 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFQDTW, LSTQAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-12 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFQDTW, LSTQAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-13 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFRDVW, IKPKSGGGTS and ATFFQGYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQALQTPYT in sequence. The SY-14 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFRDVW, IKPKSGGGTS and ATFFQGAFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQSLQTPYT in sequence. The SY-15 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSNTW, IKSQPDGGTT and ATFFQGYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQSNGYNY, LGS and MQALQTPYT in sequence. The SY-16 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFTDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-17 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFTDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-18 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDAW, IKTKALGGTS and TTLFQGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTLMNRNGYNY, LSS and MQALQTPYT in sequence. The SY-19 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFDFTNAW, IRSNPNGGTA and VTQFLGSFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHRNGRSY, LGS and MQALQPPYS in sequence. The SY-20 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTNTYG, ISIGNSYK and AKLGVGYSSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are DSNIGNNY, KNN and AAWDNSLSGVI in sequence. The SY-21 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFILSDHY, SRNKANSYTT and VRSILVTTGGNYFEC in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QGISSS, DAS and QQSKSYPLT in sequence. The SY-22 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTDYA, IDTSTGNP and VRGSAVHILRFFDWRRFDD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGSNF, DVN and CSYAASYTWV in sequence. The SY-26 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence. The SY-27 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGLRV in sequence. The SY-28 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, MQNSGHT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYNGLRV in sequence. The SY-29 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYY, VQNSGNT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-30 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYY, VQNSGNI and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVSNGHS, DTS and LLSYSGVRV in sequence. The SY-31 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYH, IQNSGNT and GRVGHDFWGGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-32 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVSRGSYF, IQNSGST and GRQGHDFWSGYLRFDYNTDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTN and LLSYSGVRV in sequence. The SY-33 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDI, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV, respectively. The SY-34 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-35 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, IQNSGST and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTS and LLSYSGVRV in sequence. The SY-36 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGNYF, MQNSGTT and GRQGHDFWSGYLRFDYNSDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGAVTSGHS, DTT and LLSYSGVRV in sequence. The SY-37 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are SDSVGRGSYF, MQNTGAT and GRMGHDFWSGYLRFDYNSDI, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are TGGVTSGHS, DTS and LLSYSGVRV, respectively. The SY-38 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTRGYA, ISGSGLST and AKDPVSPVTVVGTFFQH in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGYNY, EVT and SSYAGSNTLGV in sequence. The SY-39 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLNT and AKDPVTPITVVGTFFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYNY, EVT and SSFAGSSNLGV in sequence. The SY-40 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLST and AKDPVTPITVVGTYFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYNY, EVT and SSFAGSSNLGV in sequence. The SY-41 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFRAYA, ISGSGLNT and AKDPVTPITVVGTFFLD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SNDVGGYSY, EVT and SSFAGSSNLGV in sequence. The SY-43 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are DFSFSSHA, ISYDGGTT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QRINSRY, AAS and QQYGNSPLCS in sequence. The SY-44 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYYGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QNINSRY, AAS and QQYGNSPLCS in sequence. The SY-45 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFSFSSYA, ISYDGSNT and AKEGNAFRYFGWLPDYQYFGMDV in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSINSRY, AAS and QQYGNTPLCS in sequence. The SY-46 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VYSFKDTW, LSTEAAGATA and TTLFQGHFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQRNGYNS, LGS and MQGLQTPYT in sequence. The SY-47 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFDFNHVW, IKSKSDGATT and ATFFHAYFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are RSLLQRNGDNY, LGT and QGLQTPYT in sequence. The SY-48 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDVW, IKPKSGGGTS and ATFFQGYFDY, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLHSNGYNY, FGS and MQALQTPYT, respectively. The SY-49 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are VFSFSDAW, IKTTALGATS and TTLFGGAFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QTVMNRNGYNY, LSS and MQSVQTPYT in sequence. The SY-50 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are EFTLSRHL, ISIGSSYI and VRLGVGFTSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are NSNIGSNY, KNN and AAWDNSLSGVA in sequence. The SY-51 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFNRYG, ISIGSSYK and AKLGVGYSSSSEDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are DSNIGNNY, KNN and AAWDNSLGGVV in sequence. The SY-52 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GFTFSDHY, TRNRANSYRT and VRGPLVGKTGFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QGIRNY, TVS and LQHSSYPYS in sequence. The SY-53 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are HFTFSSYA, IWYDGTYK and AKSLTGALEG in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLNSDDGNTY, TVS and MHRVEFPRT in sequence. The SY-54 is composed of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYSFTNYA, IDTYTGNP and ARGSAVHILQFFDWRRFDD in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are SSDVGGYNF, DVS and CSYAGSYTWV in sequence. The SY-55 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GDTFSSYT, INGFLGIA and ARGDAGAVRLLYYNYAMDL, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSLLQSDGKTY, EVS and MQTIQLPLT, respectively. The SY-56 is composed of a heavy chain and a light chain paired therewith; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GGTFRSYG, IVPIFGIS and VRGGAVAGAGSAGEGIFDY in sequence, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are QSVSKN, GAS and QQHHSWPPFT in sequence. The SY-57 consists of a heavy chain and a paired light chain; the amino acid sequences of CDR1, CDR2 and CDR3 of the heavy chain are GYTFTKFG, ITPYNGNR and SRDGDRAAVSSF, respectively, and the amino acid sequences of CDR1, CDR2 and CDR3 of the light chain are ESLFFDSLNKNY, WAS and QQYIMSPWT, respectively.

3. The hepatitis B virus monoclonal neutralizing antibody composition according to claim 2, characterized in that SY The heavy chain variable region sequence with -1 is Seq ID No.1, and the light chain variable region sequence is Seq ID No.2; The heavy chain variable region sequence of SY-2 is Seq ID No. 3, and the light chain variable region sequence is Seq ID No. 4; The heavy chain variable region sequence of SY-3 is Seq ID No. 5, and the light chain variable region sequence is Seq ID No. 6; The heavy chain variable region sequence of SY-5 is Seq ID No. 7, and the light chain variable region sequence is Seq ID No. 8; The heavy chain variable region sequence of SY-6 is Seq ID No. 9, and the light chain variable region sequence is Seq ID No. 10; The heavy chain variable region sequence of SY-7 is Seq ID No. 11, and the light chain variable region sequence is Seq ID No. 12; The heavy chain variable region sequence of SY-8 is Seq ID No. 13, and the light chain variable region sequence is Seq ID No. 14; The heavy chain variable region sequence of SY-9 is Seq ID No. 15, and the light chain variable region sequence is Seq ID No. 16; The heavy chain variable region sequence of SY-10 is Seq ID No. 17, and the light chain variable region sequence is Seq ID No. 18; The heavy chain variable region sequence of SY-11 is Seq ID No. 19, and the light chain variable region sequence is Seq ID No. 20; The heavy chain variable region sequence of SY-12 is Seq ID No. 21, and the light chain variable region sequence is Seq ID No. 22; The heavy chain variable region sequence of SY-13 is Seq ID No. 23, and the light chain variable region sequence is Seq ID No. 24; The heavy chain variable region sequence of SY-14 is Seq ID No. 25, and the light chain variable region sequence is Seq ID No. 26; The heavy chain variable region sequence of SY-15 is Seq ID No. 27, and the light chain variable region sequence is Seq ID No. 28; The heavy chain variable region sequence of SY-16 is Seq ID No. 29, and the light chain variable region sequence is Seq ID No. 30; The heavy chain variable region sequence of SY-17 is Seq ID No. 31, and the light chain variable region sequence is Seq ID No. 32; The heavy chain variable region sequence of SY-18 is Seq ID No. 33, and the light chain variable region sequence is Seq ID No. 34; The heavy chain variable region sequence of SY-19 is Seq ID No. 35, and the light chain variable region sequence is Seq ID No. 36; The heavy chain variable region sequence of SY-20 is Seq ID No. 37, and the light chain variable region sequence is Seq ID No. 38; The heavy chain variable region sequence of SY-21 is Seq ID No. 39, and the light chain variable region sequence is Seq ID No. 40; The heavy chain variable region sequence of SY-22 is Seq ID No. 41, and the light chain variable region sequence is Seq ID No. 42; The heavy chain variable region sequence of SY-26 is Seq ID No. 43, and the light chain variable region sequence is Seq ID No. 44; The heavy chain variable region sequence of SY-27 is Seq ID No. 45, and the light chain variable region sequence is Seq ID No. 46; The heavy chain variable region sequence of SY-28 is Seq ID No. 47, and the light chain variable region sequence is Seq ID No. 48; The heavy chain variable region sequence of SY-29 is Seq ID No. 49, and the light chain variable region sequence is Seq ID No. 50; The heavy chain variable region sequence of SY-30 is Seq ID No. 51, and the light chain variable region sequence is Seq ID No. 52; The heavy chain variable region sequence of SY-31 is Seq ID No. 53, and the light chain variable region sequence is Seq ID No. 54; The heavy chain variable region sequence of SY-32 is Seq ID No. 55, and the light chain variable region sequence is Seq ID No. 56; The heavy chain variable region sequence of SY-33 is Seq ID No. 57, and the light chain variable region sequence is Seq ID No. 58; The heavy chain variable region sequence of SY-34 is Seq ID No. 59, and the light chain variable region sequence is Seq ID No. 60; The heavy chain variable region sequence of SY-35 is Seq ID No. 61, and the light chain variable region sequence is Seq ID No. 62; The heavy chain variable region sequence of SY-36 is Seq ID No. 63, and the light chain variable region sequence is Seq ID No. 64; The heavy chain variable region sequence of SY-37 is Seq ID No. 65, and the light chain variable region sequence is Seq ID No. 66; The heavy chain variable region sequence of SY-38 is Seq ID No. 67, and the light chain variable region sequence is Seq ID No.

68. The heavy chain variable region sequence of SY-39 is Seq ID No. 69, and the light chain variable region sequence is Seq ID No. 70; The heavy chain variable region sequence of SY-40 is Seq ID No. 71, and the light chain variable region sequence is Seq ID No. 72; The heavy chain variable region sequence of SY-41 is Seq ID No. 73, and the light chain variable region sequence is Seq ID No. 74; The heavy chain variable region sequence of SY-43 is Seq ID No. 75, and the light chain variable region sequence is Seq ID No. 76; The heavy chain variable region sequence of SY-44 is Seq ID No. 77, and the light chain variable region sequence is Seq ID No. 78; The heavy chain variable region sequence of SY-45 is Seq ID No. 79, and the light chain variable region sequence is Seq ID No. 80; The heavy chain variable region sequence of SY-46 is Seq ID No. 81, and the light chain variable region sequence is Seq ID No. 82; The heavy chain variable region sequence of SY-47 is Seq ID No. 83, and the light chain variable region sequence is Seq ID No. 84; The heavy chain variable region sequence of SY-48 is Seq ID No. 85, and the light chain variable region sequence is Seq ID No. 86; The heavy chain variable region sequence of SY-49 is Seq ID No. 87, and the light chain variable region sequence is Seq ID No. 88; The heavy chain variable region sequence of SY-50 is Seq ID No. 89, and the light chain variable region sequence is Seq ID No. 90; The heavy chain variable region sequence of SY-51 is Seq ID No. 91, and the light chain variable region sequence is Seq ID No. 92; The heavy chain variable region sequence of SY-52 is Seq ID No. 93, and the light chain variable region sequence is Seq ID No. 94; The heavy chain variable region sequence of SY-53 is Seq ID No. 95, and the light chain variable region sequence is Seq ID No. 96; The heavy chain variable region sequence of SY-54 is Seq ID No. 97, and the light chain variable region sequence is Seq ID No. 98; The heavy chain variable region sequence of SY-55 is Seq ID No. 99, and the light chain variable region sequence is Seq ID No. 100; The heavy chain variable region sequence of SY-56 is Seq ID No. 101, and the light chain variable region sequence is Seq ID No. 102; The heavy chain variable region sequence of SY-57 is Seq ID No. 103, and the light chain variable region sequence is Seq ID No.

104.

4. A biomaterial, characterized in that, The biomaterial is selected from one of the following (A)-(B): (A) A nucleic acid molecule encoding the monoclonal antibody against hepatitis B virus HBsAg as described in claim 1, or the monoclonal antibody composition against hepatitis B virus HBsAg as described in claim 3; (B) Recombinant plasmids and recombinant cells containing the nucleic acid molecules described in (A).

5. The biomaterial according to claim 4, characterized in that, The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 1 is shown in Seq ID No. 105, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 2 is shown in Seq ID No.

106. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 3 is shown in Seq ID No. 107, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 4 is shown in Seq ID No.

108. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 5 is shown in Seq ID No. 109, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 6 is shown in Seq ID No.

110. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 7 is shown in Seq ID No. 111, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 8 is shown in Seq ID No.

112. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 9 is shown in Seq ID No. 113, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 10 is shown in Seq ID No.

114. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 11 is shown in Seq ID No. 115, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 12 is shown in Seq ID No.

116. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 13 is shown in Seq ID No. 117, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 14 is shown in Seq ID No.

118. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 15 is shown in Seq ID No. 119, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 16 is shown in Seq ID No.

120. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 17 is shown in Seq ID No. 121, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 18 is shown in Seq ID No.

122. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 19 is shown in Seq ID No. 123, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 20 is shown in Seq ID No.

124. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 21 is shown in Seq ID No. 125, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 22 is shown in Seq ID No.

126. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 23 is shown in Seq ID No. 127, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 24 is shown in Seq ID No.

128. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 25 is shown in Seq ID No. 129, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 26 is shown in Seq ID No.

130. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 27 is shown in Seq ID No. 131, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 28 is shown in Seq ID No.

132. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 29 is shown in Seq ID No. 133, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 30 is shown in Seq ID No.

134. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 31 is shown in Seq ID No. 135, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 32 is shown in Seq ID No.

136. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 33 is shown in Seq ID No. 137, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 34 is shown in Seq ID No.

138. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 35 is shown in Seq ID No. 139, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 36 is shown in Seq ID No.

140. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 37 is shown in Seq ID No. 141, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 38 is shown in Seq ID No.

142. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 39 is shown in Seq ID No. 143, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 40 is shown in Seq ID No.

144. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 41 is shown in Seq ID No. 145, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 42 is shown in Seq ID No.

146. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 43 is shown in Seq ID No. 147, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 44 is shown in Seq ID No.

148. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 45 is shown in Seq ID No. 149, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 46 is shown in Seq ID No.

150. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 47 is shown in Seq ID No. 151, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 48 is shown in Seq ID No.

152. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 49 is shown in Seq ID No. 153, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 50 is shown in Seq ID No.

154. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 51 is shown in Seq ID No. 155, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 52 is shown in Seq ID No.

156. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 53 is shown in Seq ID No. 157, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 54 is shown in Seq ID No.

158. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 55 is shown in Seq ID No. 159, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 56 is shown in Seq ID No.

160. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 57 is shown in Seq ID No. 161, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 58 is shown in Seq ID No.

162. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 59 is shown in Seq ID No. 163, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 60 is shown in Seq ID No.

164. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 61 is shown in Seq ID No. 165, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 62 is shown in Seq ID No.

166. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 63 is shown in Seq ID No. 167, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 64 is shown in Seq ID No.

168. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 65 is shown in Seq ID No. 169, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 66 is shown in Seq ID No.

170. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 67 is shown in Seq ID No. 171, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 68 is shown in Seq ID No.

172. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 69 is shown in Seq ID No. 173, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 70 is shown in Seq ID No.

174. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 71 is shown in Seq ID No. 175, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 72 is shown in Seq ID No.

176. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 73 is shown in Seq ID No. 177, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 74 is shown in Seq ID No.

178. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 75 is shown in Seq ID No. 179, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 76 is shown in Seq ID No.

180. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 77 is shown in Seq ID No. 181, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 78 is shown in Seq ID No.

182. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 79 is shown in Seq ID No. 183, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 80 is shown in Seq ID No.

184. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 81 is shown in Seq ID No. 185, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 82 is shown in Seq ID No.

186. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 83 is shown in Seq ID No. 187, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 84 is shown in Seq ID No.

188. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 85 is shown in Seq ID No. 189, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 86 is shown in Seq ID No.

190. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 87 is shown in Seq ID No. 191, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 88 is shown in Seq ID No.

192. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 89 is shown in Seq ID No. 193, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 90 is shown in Seq ID No.

194. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 91 is shown in Seq ID No. 195, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 92 is shown in Seq ID No.

196. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 93 is shown in Seq ID No. 197, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 94 is shown in Seq ID No.

198. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 95 is shown in Seq ID No. 199, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 96 is shown in Seq ID No.

200. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 97 is shown in Seq ID No. 201, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 98 is shown in Seq ID No.

202. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 99 is shown in Seq ID No. 203, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 100 is shown in Seq ID No.

204. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 101 is shown in Seq ID No. 205, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 102 is shown in Seq ID No.

206. The nucleic acid sequence encoding the heavy chain variable region shown in Seq ID No. 103 is shown in Seq ID No. 207, and the nucleic acid sequence encoding the light chain variable region shown in Seq ID No. 104 is shown in Seq ID No.

208.

6. The method for preparing the monoclonal antibody against hepatitis B virus HBsAg as described in claim 1, characterized in that, include: The recombinant cells described in claim 4 are used to express the recombinant plasmid described in claim 4, and the expressed protein is extracted and purified to obtain the monoclonal antibody against hepatitis B virus HBsAg.

7. The use of the monoclonal antibody against hepatitis B virus HBsAg as described in claim 1, or the monoclonal antibody composition against hepatitis B virus HBsAg as described in any one of claims 2-3, or the biological material as described in claim 4, in the preparation of hepatitis B virus prevention and treatment drugs or hepatitis B virus detection reagents.

8. A drug for the prevention and treatment of hepatitis B virus, characterized in that, The composition contains the monoclonal antibody against hepatitis B virus HBsAg as described in claim 1, or the monoclonal antibody composition against hepatitis B virus HBsAg as described in any one of claims 2-3.

9. A hepatitis B virus detection reagent, characterized in that, The composition contains the monoclonal antibody against hepatitis B virus HBsAg as described in claim 1, or the monoclonal antibody composition against hepatitis B virus HBsAg as described in any one of claims 2-3.

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