Antibody screening method based on heavy chain library/light chain library of infectable virus particle type antibody and preparation method of heavy chain library/light chain library

Abstract

The invention relates to a novel antibody screening method, the method comprises the following steps: (1) anti-body library construction: constructing a heavy chain library of infectable virus particle type antibody and a light chain library of infectable virus particle type antibody; (2) a first screening step: infecting animal cells with the heavy chain library of infectable virus particle type antibody and the light chain library of infectable virus particle type antibody, selecting the animal cells with a specific antibody expression ability by using a specific antigen so as to obtain positive cell strains; (3) a second screening step: obtaining the cDNA sequences of the heavy chain variable region and the light chain variable region of the specific antibody from the positive cell strains, inoculating light chain signal peptide sequence of the specific antibody, inserting an expression vector, and constructing an expression cell strain of the specific antibody so as to prepare the specific antibody through expression. The screening method has the advantages of simple and high efficient technology and practicality. The gene engineering antibody which is obtained through screening has a high specificity and high affinity.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals and relates to the technical field of antibody screening. More specifically, the present invention relates to the establishment of an infectable virus particle type antibody heavy chain library and an infectable virus particle type antibody light chain library and a novel antibody screening method. Background technique [0002] Antibody molecules are the most widely used protein molecules in the field of biomedicine, and therapeutic antibodies and diagnostic antibodies have broad application prospects and huge market value. Since the first genetically engineered antibody human-mouse chimeric antibody was born in 1984, new genetically engineered antibodies such as humanized antibodies, monovalent small molecule antibodies, and multivalent small molecule antibodies have emerged continuously. [0003] Hybridoma cell technology has brought revolutionary changes to the production technology...

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to address the defects that the hybridoma cell technology in the existing antibody screening technology cannot prepare specific antibodies and humanized antibodies against rare antigens; for various antibody screening technologies based on antibody libraries There are defects such as low antigen affinity and poor specificity; in view of the defects that mammalian cell display technology cannot prepare single-chain antibodies, an antibody heavy chain library that can infect virus particles and an antibody light chain library that can infect virus particles , using the antibody library to establish a new antibody screening method to screen humanized antibodies with high specificity and high affinity that are closer to in vivo conditions

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