UNIT 3.2 HIV Protease Assays
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Pharmacology
How to Cite
Furfine, E. S. 2001. HIV Protease Assays. Current Protocols in Pharmacology. 00:3.2:3.2.1–3.2.12.
- Published Online: 1 MAY 2001
- Published Print: MAR 1998
The fluorescence-based assay described in this unit is used not only to screen large numbers of compounds in a 96-well format for the ability to inhibit human immunodeficiency virus-1 (HIV) protease, but also to determine accurately the affinity of inhibitors for the enzyme. A method for performing an active site titration, using tight-binding inhibitors, and determining the Km of a substrate is also provided. Because numerous potent inhibitors of HIV protease have been identified, information is provided on the analysis required to deal with these potent compounds and with fluorometric enzyme assay data in general.