A HIGH THROUGHPUT ASSAY FOR INHIBITORS OF HIV-1 PROTEASE - SCREENING OF MICROBIAL METABOLITES

被引：19

作者：

SARUBBI, E ^{[1
]}

NOLLI, ML ^{[1
]}

ANDRONICO, F ^{[1
]}

STELLA, S ^{[1
]}

SADDLER, G ^{[1
]}

SELVA, E ^{[1
]}

SICCARDI, A ^{[1
]}

DENARO, M ^{[1
]}

机构：

[1] UNIV MILAN,DIPARTIMENTO BIOL & GENET,I-20122 MILAN,ITALY

来源：

FEBS LETTERS | 1991年 / 279卷 / 02期

关键词：

HUMAN IMMUNODEFICIENCY VIRUS; PROTEINASE; FUSION PROTEIN; IMMUNOLOGICAL ASSAY; MICROBIAL ALKALINE PROTEASE INHIBITOR;

D O I：

10.1016/0014-5793(91)80164-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel method for discovery of HIV-1 protease inhibitors in complex biological samples has been developed. The assay is based on two specific reagents: a recombinant protein constituted by a portion of the HIV-1 Gag polyprotein comprising the p17-p24 cleavage site, fused to E. coli beta-galactosidase, and a monoclonal antibody which binds the fusion protein in the Gag region. Binding occurs only if the fusion protein has not been cleaved by the HIV-1 protease. The assay has been adapted for the screening of large numbers of samples in standard 96-well microtiter plates. Using this method about 12000 microbial fermentation broths have been tested and several HIV-1 protease inhibitory activities have been detected. One of these has been studied in detail.

引用

页码：265 / 269

页数：5

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