Antigenic Epitope Analysis and Preparation of Monoclonal Antibody of MMP-9

doi:10.13560/j.cnki.biotech.bull.1985.2018-1059

Abstract

Abstract: In this study,we used bioinformatics analysis to predict antigenic epitope region of MMP-9,which was designated as Δmmp9. Analyzing the nucleotide sequence of the Δmmp9,the primers were designed to amplify the target fragment,and the recombinant pET28α（+）-Δmmp9 expression vector was constructed,and transferred into Escherichia coli competent cell BL21（DE3）for inducing the recombinant protein Δmmp9. Then recombinant protein Δmmp9 was purified,and then was used to immunize BALB/c mice as antigen. Furthermore,7 hybridoma cell lines steadily secreting monoclonal antibodies（MAbs）were screened,and titers of seven MAbs all reached 1：240 000. In summary,MMP-9 antigenic epitope region is screened by bioinformatics methods and its MAbs are prepared successfully,its promising specificity is confirmed by antibody pairing test,and 3 captured antibodies and corresponding labeled antibodies are screened out.

Key words: MMP-9, bioinformatics, epitope, monoclonalantibody

XU Ying-xue, YIN Huan-cai. Antigenic Epitope Analysis and Preparation of Monoclonal Antibody of MMP-9[J]. Biotechnology Bulletin, 2019, 35(7): 134-140.

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