HIV-1嵌合抗原的纯化及免疫原性分析

Purification and Analysis of Antigencity of Chimeric Antigen of HIV-1

为获得高效的H IV诊断试剂,选定H IV-1的外膜蛋白env中469-511 aa,538-674 aa和700-734 aa 3处包含较多抗原位点的区域作为免疫抗原,用PCR的方法从H IV-1全基因扩增编码这3处片段的基因序列,将它们克隆到原核表达载体中,利用大肠杆菌表达系统表达嵌合蛋白.结果发现,3段嵌合基因能在大肠杆菌BL21(Star)中表达,通过N i-sepharose 4B金属N i螯合层析柱分离纯化目的产物,酶联免疫检测结果表明,纯化抗原有较强的抗原特异性.

In order to get the high-efficient diagnosing reagent of HIV, we choose three segments of HIV-1 envelope protein as the immune antigen, 469-511 aa, 538-674aa and 700-734aa, which include more epitope. These three segment genes were obtained by PCR amplification and cloned into prokaryotic expressing vector. The results show that chimeric env protein can be expressed in E. coli BL21 strain high efficiently. We purified the protein by metal Ni-sepharose 4B, the data of ELISA demonstrate that purified chimeric antigen has good antigenic specificity.