AT-2灭活HIV-1病毒及纯化回收鉴定

AT-2-Inactivation of HIV-1 Virus and Its Purification and Identification

目的制备具备完整空间构型且纯度在90%以上的灭活HIV-1病毒,用于HIV疫苗研究。方法应用化学制剂2,2’-dithiodipyridine(aldrithiol-2;AT-2)灭活HIV-1病毒,对灭活的病毒超速离心浓缩并洗涤去除灭活用的化学制剂AT-2。采用分子筛技术去除灭活病毒中残存的杂质蛋白质。结果 250μmol/L AT-2与HIV-137℃作用1 h可以彻底灭活病毒的感染性,同时保留病毒的免疫原性。灭活的病毒纯化后检测不到AT-2的残留,检测牛血清蛋白残余量低于50 ng/mL。结论灭活的HIV-1病毒经过纯化后纯度达到95.6%,可以满足作为HIV疫苗研究的免疫刺激剂的使用要求。

Objective To prepare an inactivated HIV-1 virus with configurational integrity and more than 90% purity for HIV vaccine research.Methods HIV-1 virus was inactivated by 2,2’-dithiodipyridine（aldrithiol-2,AT-2） with complete immunogenicity.Ultracentrifugation and washing was conducted to remove the chemical reagent used for virus inactivation.The residual impurity proteins in the inactivated virus was removed by molecular sieve technology.Results For chemical inactivation,HIV-1 virus was treated with 250 μM AT-2 for 1 h at 37℃.Chemically inactivated noninfectious SIV with conformationally and functionally intact envelope glycoproteins was produced by treatment with AT-2 as described elsewhere[1-2].No AT-2 residue was detected in the purified inactivated virus.Residual bovine serum albumin was detected less than 50 ng /mL.Conclusions In this study HIV-1 virus was successfully inactivated with AT-2 and reached a purity of 95.6%.It can meet the requirements as an immune stimulating agent in HIV vaccine research.