[Analysis on bioactivity of HIV-1 integrase by ELISA method].
To develop an ELISA-based method for analyzing biologic activities of HIV-1 integrase and for high throughput screening of integrase inhibitors.



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.



(1) The specific activity of the purified integrase was 54.92 units/mg of protein. (2)IC(50) (concentration causing 50% inhibition of integrase) of luffin-a was (0.63 +/- 0.026) micromol/L.



The non-radioactive assay can be used for analysis of bioactivities and high throughput screening of inhibitors of HIV-1 integrase.
Feng
Wei-Hong
WH
Department of Biochemistry and Molecular Biology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
Huang
Jian-Song
JS
Zhan
Jin-Biao
JB
D002384:Catalysis / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008163:Luminescent Measurements; D054789:Ribosome Inactivating Proteins, Type 1 / Q000494:pharmacology; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D054789:Ribosome Inactivating Proteins, Type 1; C063258:luffin-a protein, Luffa cylindrica; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1