MMP-9蛋白的抗原表位分析及单克隆抗体制备  

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

作　　者：许映雪 尹焕才[1] XU Ying-xue;YIN Huan-cai(Suzhou Institute of Biomedical Engineering Technology,Chinese Academy of Sciences,Suzhou 215163;University of Chinese Academy of Sciences,Beijing 100049)

机构地区：[1]中国科学院苏州生物医学工程技术研究所,苏州215163 [2]中国科学院大学,北京100049

出　　处：《生物技术通报》2019年第7期134-140,共7页Biotechnology Bulletin

基　　金：医疗器械灭菌效果评价系统的产业化(2018SYHZ0020)

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。In this study,we used bioinformatics analysis to predict antigenic epitope region of MMP-9,which was designated asΔmmp9.Analyzing the nucleotide sequence of theΔmmp9,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 proteinΔmmp9.Then recombinant proteinΔmmp9 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.

关 键 词：MMP-9 生物信息学 抗原表位 单克隆抗体 

分 类 号：R73[医药卫生—肿瘤]