Method for analyzing epitope of monoclonal antibody by using yeast surface display system and application of method in vaccine development

Abstract

The invention discloses a method for analyzing epitope of monoclonal antibody by using a yeast surface display system and application of the method in vaccine development. The method comprises the following steps of: (1) constructing a DNA library of target protein; (2) displaying the DNA library on the surface of yeast to obtain a yeast display library; (3) mixing the monoclonal antibody of the target protein and a yeast display library A, and screening yeast combined with the monoclonal antibody; and (4) extracting plasmid of the screened yeast, sequencing, and analyzing the epitope of the target protein. Compared with the traditional method, the method has the advantages that: (1) ways of modifying the protein by yeast cells on are more, and high molecular weight and complicated protein can be displayed; (2) the yeast can be screened one by one by fluorescence-activated cell sorting (FACS), and the accuracy and screening flux are improved; (3) the yeast can independently complete self growing and reproducing process, the operation is simple and convenient and interference factors are a few; and (4) the yeast is an immunologic adjuvant, and can check whether the epitope can induce the generation of antibody with similar activity again.

Description

technical field [0001] The invention relates to a method for analyzing the antigenic epitope of monoclonal antibody by using yeast surface display system and its application in vaccine development. Background technique [0002] The host's antibody response plays an important role in the process of resisting pathogenic infection, and the polyclonal antibody response in the body is composed of monoclonal antibodies against different antigens or different epitopes of the same antigen. A deep understanding of this complex process can provide key information for the study of disease pathogenesis, and can also provide an important theoretical basis for the development of efficient vaccines and disease treatment. [0003] Among the many research methods of antigenic epitopes, the phage display peptide library technology emerged in 1990 (Scott, J.K. and G.P.Smith, Searching for peptide ligands with an epitope library. Science, 1990.249(4967): p.386-90. ), that is, using a randomly ...

AI Technical Summary

Problems solved by technology

However, the phage display peptide library technology also has great limitations: (1) Most of the short peptide libraries that can be generated are linear epitopes, which cannot well mimic the conformational epitopes that account for more than 80% of antigenic epitopes; (2) The foreign molecules that can be accommodated by the capsid protein of phage are small and simple in structure, and lack of modification of foreign proteins. Many cytotoxic molecules and eukaryotic proteins in this system are difficult to express and display; (3) The individual phage is too small , mostly using affinity chromatography columns or affinity polystyrene plates for screening, the screening efficiency is low; (4) After each round of screening, the phage must re-infect the host cells in order to obtain enough virus particles for the next round of screening, recombinant virus The difference in infection ability between particles will result in certain amplification advantages; (5) phages are easily spread by air, thus easily causing cross-contamination

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