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.
作者
许映雪
尹焕才
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)
出处
《生物技术通报》
CAS
CSCD
北大核心
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)
