UNIT 3.2 HIV Protease Assays

  1. Eric S. Furfine

Published Online: 1 MAY 2001

DOI: 10.1002/0471141755.ph0302s00

Current Protocols in Pharmacology

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.

Author Information

  1. Glaxo Wellcome Research, Research Triangle Park, North Carolina

Publication History

  1. Published Online: 1 MAY 2001
  2. 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.