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hepatitis B surface antigen resistant antibody and application thereof

一种抗体、抗原的技术，应用在抗体、应用、抗病毒剂等方向

Active Publication Date: 2017-04-19
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no clear and effective active immunotherapy drug / vaccine for the treatment of chronic HBV infection

Method used

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  • hepatitis B surface antigen resistant antibody and application thereof
  • hepatitis B surface antigen resistant antibody and application thereof
  • hepatitis B surface antigen resistant antibody and application thereof

Examples

Experimental program
Comparison scheme
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specific Embodiment approach

[1732] The invention will now be described with reference to the following examples, which are intended to illustrate the invention, but not to limit it.

[1733] Unless otherwise specified, the molecular biology experiment methods and immunoassay methods used in the present invention are basically with reference to J.Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, and F.M.Ausubel et al., Molecular Biology Experimental Guide, 3rd Edition, John Wiley & Sons, Inc., 1995. The method described in; restriction endonucleases were used in accordance with the conditions recommended by the product manufacturer. Those skilled in the art understand that the examples describe the present invention by way of example and are not intended to limit the scope of the claimed invention.

Embodiment 1

[1734] Example 1: Mouse monoclonal antibody 6D11 specifically binding to HBsAg and its humanization

[1735] 1.1: Characterization of mouse monoclonal antibody 6D11

[1736] The mouse monoclonal antibody 6D11 (hereinafter also referred to as 6D11-mAb or mAb for short) that specifically binds to HBsAg has been prepared according to conventional immunological methods. The amino acid sequence of the heavy chain variable region of the mouse monoclonal antibody 6D11 is shown in SEQ ID NO:1, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:2.

[1737] SEQ ID NO: 1

[1738] DVQLQESGPGLVKPSQSLSLTCSVTGYPITSGYHWNWIRQFPGNKLVWMGYISYDGSDHYNPSLENRISITRDISKNQFFLILRSVTTEDTGKYFCASGFDHWGQGTTLTVSS

[1739] SEQ ID NO: 2

[1740]DVVMTQTPLSLPVSLGDQASISCRSSQSLVHSYGDTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVETEDLGVYFCSQNTHVPYTFGGGTKLEIKR

[1741] Further, the method described by Kabat et al. (Kabat et al., Sequences of Proteins of Immunological Interest...

Embodiment 2

[1760] Example 2: Single-point substitution of amino acids in the CDR region of humanized antibody B-S3-45

[1761] The amino acids at each site of the six CDR regions of the humanized antibody B-S3-45 were substituted with 20 naturally occurring amino acids at a single point. Using the cloning methods in Examples 1.3 and 1.4, the recombinant vector expressing the phage antibody was obtained, wherein the phage antibody had a single point mutation in the CDR region compared with B-S3-45. Amino acid substitutions at the mutation site were introduced using degenerate primers.

[1762] Taking the first amino acid of the heavy chain variable region CDR3 (HCDR3) as an example, oligonucleotide primers H3R1 and B45-H3F were designed (the sequences of which are shown in Table 5). For the annealing position on the coding gene, see Figure 4 . The specific protocol of PCR is as follows: using the coding gene of B-S3-45 as a template, primer pairs B45-VhF / H3R1 and B45-H3F / B45-VkR were ...

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Abstract

The invention relates to a hepatitis B surface antigen (HBsAg) resistant antibody (in particular to a humanized antibody), nucleic acid molecules for coding the antibody, a method for preparing the antibody and a pharmaceutical composition containing the antibody. In addition, the invention relates to application of the antibody and the pharmaceutical composition. The antibody and the pharmaceutical composition can be used for preventing and / or treating HBV infection and diseases (such as hepatitis B) related to the HBV infection, used for neutralizing the HBV virulence in the body of a subject (such as a person), or used for reducing the HBV DNA and / or HBsAg serum level in the body of the subject.

Description

technical field [0001] The invention relates to the field of molecular virology and immunology, especially the field of treating hepatitis B virus (Hepatitis Bvirus, HBV) infection. Specifically, the present invention relates to antibodies (especially humanized antibodies) against hepatitis B surface antigen (HBsAg), nucleic acid molecules encoding them, methods for preparing them, and pharmaceutical compositions containing them. The pharmaceutical composition can be used to prevent and / or treat HBV infection or diseases related to HBV infection (such as hepatitis B), to neutralize the virulence of HBV in a subject (such as a human), or to treat Reduce the serum levels of HBV DNA and / or HBsAg in patients. Therefore, the present invention further relates to the use of said antibodies (especially humanized antibodies) and variants thereof in the preparation of pharmaceutical compositions for preventing and / or treating HBV infection or related to HBV infection diseases (such as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08C12N15/13C12P21/00A61K39/42A61P31/20
CPCC07K16/082A61K2039/505C07K2317/76C07K2317/92C07K2317/56C07K2317/565C07K2317/24C07K2317/622C07K2317/94A61P31/20C07K16/10C07K2319/61
Inventor 罗文新周兵张娟袁权张天英张军夏宁邵
Owner XIAMEN UNIV
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