title: A high throughput assay for inhibitors of HIV-1 protease. Screening of microbial metabolites.
abstract: 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.
authors.affiliation: Lepetit Research Center, MMDRI, Gerenzano VA, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Sarubbi
authors.forename: M L
authors.initials: ML
authors.lastname: Nolli
authors.forename: F
authors.initials: F
authors.lastname: Andronico
authors.forename: S
authors.initials: S
authors.lastname: Stella
authors.forename: G
authors.initials: G
authors.lastname: Saddler
authors.forename: E
authors.initials: E
authors.lastname: Selva
authors.forename: A
authors.initials: A
authors.lastname: Siccardi
authors.forename: M
authors.initials: M
authors.lastname: Denaro
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D015683:Gene Products, gag / Q000378:metabolism; D017320:HIV Protease Inhibitors; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D015683:Gene Products, gag; D017320:HIV Protease Inhibitors; D011480:Protease Inhibitors; D011993:Recombinant Fusion Proteins