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ELISA法分析HIV-1整合酶的生物学活性 被引量：3

Analysis on bioactivity of HIV-1 integrase by ELISA method
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摘要 目的:建立一种能够较好地分析HIV-1整合酶的生物学活性以及大规模地筛选整合酶抑制剂的方法。方法:HIV-1整合酶经表达复性及纯化后,采用一种基于Biotin-Avidin EILSA(BA-ELISA)的方法,使用化学发光底物测定分析整合酶的生物学活性,以及型核糖体失活蛋白luffin-a对整合酶的抑制作用。结果:1采用BA-ELISA法得到整合酶的比活性为54.92U/mg蛋白;2luffin-a对整合酶的半数抑制浓度为(0.63±0.026)μmol/L。结论:本实验所采用的基于BA-ELISA的方法能够较好地分析HIV-1整合酶的生物活性,适合于大规模体外HIV-1整合酶抑制剂的筛选。 Objective： To develop an ELISA-based method for analyzing biologic activities of HIV-1 integrase and for high throughput screening of integrase inhibitors. Methods： After expression, renaturation and purification of integrase, the bioactivity of integrase and the inhibition of luffin-a were evaluated with an in vitro assay based on biotin-avidin EILSA and chemiluminescent substrates. Results： （1）The specific activity of the purified integrase was 54.92 units/mg of protein. （2）ICs0（concentration causing 50% inhibition of integrase） of luffin-a was （0. 63±0. 026）μmol/L. Conclusion： The non-radioactive assay can be used for analysis of bioactivities and high throughput screening of inhibitors of HIV-1 integrase.
作者 冯微宏 黄建松 詹金彪
机构地区 浙江大学医学院生物化学与分子生物学教研室
出处 《浙江大学学报（医学版）》 CAS CSCD 2007年第2期179-184,共6页 Journal of Zhejiang University(Medical Sciences)
基金 国家自然科学基金(30670424)资助项目
关键词 整合酶类/遗传学 人类免疫缺陷Ⅰ型病毒 生物活性 酶联免疫吸附测定 Integrases/genet Human immunodeficiency virus type 1 Bioactivity Enzyme -linked immunosorbent assay
分类号 R446.6 [医药卫生—诊断学]
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参考文献14

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浙江大学学报（医学版）
2007年 第2期

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