       Document 0320
 DOCN  M9640320
 TI    Automation of a high-performance liquid chromatography-based enzyme
       assay: evaluation of inhibition constants for human immunodeficiency
       virus-1 protease inhibitors.
 DT    9604
 AU    Pazhanisamy S; Stuver CM; Livingston DJ; Vertex Pharmaceuticals
       Incorporated, Cambridge, Massachusetts; 02139, USA.
 SO    Anal Biochem. 1995 Jul 20;229(1):48-53. Unique Identifier : AIDSLINE
       MED/96140929
 AB    Enzyme-based assays are commonly employed in clinical and pharmaceutical
       laboratories to aid in quantitation of organic substances. Many enzyme
       assays are tedious, requiring the addition of reagents at multiple time
       intervals. The HPLC-based analysis of reaction products requires an
       additional step of vialing the samples and placing them in the
       autosampler. Such time-consuming, repetitive procedures are ideally
       suited for automation. We automated an HIV protease assay for the
       purposes of screening compounds as inhibitors of HIV protease and
       determining inhibition constants. Automation was accomplished by
       interfacing a robotic sample processor from a Gilson Model 232/401
       biocompatible automatic sample processor and injector, with a Hewlett
       Packard HPLC. We used this configuration to automate the following
       steps: (a) preparation of serial dilutions of inhibitor, (b) enzyme
       assay setup, and (c) quantitation of products of enzyme assays. The
       resulting automated method produced inhibition constants that were of
       comparable accuracy and precision to those determined by manual methods.
 DE    Amino Acid Sequence  Automation  Chromatography, High Pressure
       Liquid/*METHODS/STATISTICS & NUMER  DATA  Evaluation Studies  Human  HIV
       Protease/METABOLISM  HIV Protease Inhibitors/*PHARMACOLOGY  HIV-1/*DRUG
       EFFECTS/*ENZYMOLOGY  Kinetics  Molecular Sequence Data
       Oligopeptides/CHEMISTRY  Pepstatins/PHARMACOLOGY  Reproducibility of
       Results  Substrate Specificity  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

