MMP-9蛋白的抗原表位分析及单克隆抗体制备

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

摘要/Abstract

摘要： 利用生物信息学方法分析并预测MMP-9蛋白的抗原表位区,命名为Δmmp9。通过对Δmmp9的核苷酸序列进行分析,设计引物扩增目的片段,构建重组表达载体pET28α（+）-Δmmp9,并将其转化到BL21（DE3）中,诱导表达重组蛋白Δmmp9。纯化重组蛋白后,免疫BALB/c小鼠,筛选获得7株稳定分泌单克隆抗体的杂交瘤细胞株,检测抗体效价均达1：240 000。利用生物信息学筛选抗原表位区,成功制备单克隆抗体,通过抗体配对实验,证明其有良好特异性,并筛选出3株捕获抗体及对应标记抗体。

关键词: MMP-9, 生物信息学, 抗原表位, 单克隆抗体

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

许映雪, 尹焕才. MMP-9蛋白的抗原表位分析及单克隆抗体制备[J]. 生物技术通报, 2019, 35(7): 134-140.

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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