Analysis on bioactivity of HIV-1 integrase by ELISA method.
- Author:
Wei-Hong FENG
1
;
Jian-Song HUANG
;
Jin-Biao ZHAN
Author Information
- Publication Type:Journal Article
- MeSH: Catalysis; drug effects; Dose-Response Relationship, Drug; Enzyme Inhibitors; pharmacology; Enzyme-Linked Immunosorbent Assay; methods; HIV Integrase; chemistry; metabolism; Humans; Kinetics; Luminescent Measurements; Ribosome Inactivating Proteins, Type 1; pharmacology; Substrate Specificity
- From: Journal of Zhejiang University. Medical sciences 2007;36(2):179-184
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop an ELISA-based method for analyzing biologic activities of HIV-1 integrase and for high throughput screening of integrase inhibitors.
METHODSAfter 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.
RESULT(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.
CONCLUSIONThe non-radioactive assay can be used for analysis of bioactivities and high throughput screening of inhibitors of HIV-1 integrase.
