SCALE-UP PURIFICATION OF HIV-1 AND HIV-2 RECOMBINANT ENV POLYPEPTIDES

HIV-1 gp 41的两种高免疫原性重组Env多肽 （蛋白566）和HIV-2 gp 35（蛋白996）跨膜糖蛋白 在E.大肠杆菌的数量，纯化毫克数量 至接近同质，并用于免疫测定（蛋白质印迹和斑点印迹） 针对来自中非和西非、墨西哥和美国的血清 区分HIV-1和HIV-2感染。 此外，重组HTLV-I（rpBI）和HTLV-II（rpBII）gp 46 Env 多肽和HTLV-I p21 E Env多肽（蛋白400），先前 在大肠杆菌中表达大肠杆菌，也用于免疫测定（Western印迹）， 区分HTLV-I和HTLV-II感染。 此外，抗体 开发HTLV-I（rpBI）和HTLVII（rpBII）Env多肽， 用于抗体依赖性细胞毒性（ADCC）测定， 描绘了存在于细胞表面的类型特异性表位， HTLV-I和HTLV-II Env糖蛋白。 目前可用的血清学检测不能区分HIV-1 和HIV-2或HTLV-I和HTLV-II感染，因为显著的 抗原抗体交叉反应性，由于密切的遗传相关性 在HIV-1/HIV-2和HTLVI/HTLV-II病毒组之间。 因此 这些高免疫原性HIV-1（蛋白质566）、HIV-2 （蛋白996）、HTLV-I（rpBI和蛋白400）和HTLV-II（rpBII） 候选抗原在发展用于区分 单一以及双重病毒感染，以及作为潜在的亚单位 疫苗，需要进一步的血清学和生物学特性。

Two highly-immunogenic recombinant Env polypeptides of the HIV-1 gp41 (protein 566) and HIV-2 gp35 (protein 996) transmembrane glycoproteins were expressed in E. coli in quantity, purified in milligram quantities to near homogeneity, and used in immunoassays (Western and dot blots) against sera from Central and West Africa, Mexico, and the United States to distinguish between HIV-1 and HIV-2 infections. In addition, recombinant HTLV-I (rpBI) and HTLV-II (rpBII) gp46 Env polypeptides, and HTLV-I p2lE Env polypeptides (protein 400), previously expressed in E. coli, were also used in immunoassays (Western blot) to distinguish between HTLV-I and HTLV-ll infections. Moreover, antibodies developed to the HTLV-I (rpBI) and HTLVII (rpBII) Env polypeptides were used in antibody-dependent cellular cytotoxicity (ADCC) assays to delineate the type-specific epitopes residing on the surfaces of the HTLV-I and HTLV-II Env glycoproteins. Currently available serological tests do not distinguish between HIV-1 and HIV-2 or HTLV-I and HTLV-ll infections because of significant antigen-antibody crossreactivity due to the close genetic relatedness between HIV-l/HIV-2 and HTLVI/HTLV-II virus groups. Thus, the application of these highly-immunogenic HIV-1 (protein 566), HIV-2 (protein 996), HTLV-I (rpBI and protein 400) and HTLV-II (rpBII) candidate antigens in development of immunoassays for differentiating single, as well as dual viral infections, and as potential subunit vaccines, requires further serological and biological characterization.