Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

A fluorometric assay for HIV-protease activity using high-performance liquid chromatography.

P P Tamburini1, R N Dreyer, J Hansen

  • 1Molecular Therapeutics, Inc., West Haven, Connecticut 06516.

Analytical Biochemistry
|May 1, 1990
PubMed
Summary

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Revaccination after an Adverse Event: A Patient Uses Bayesian Reasoning to Weigh COVID and Vaccine Risks.

Medical decision making : an international journal of the Society for Medical Decision Making·2026
Same author

Long-Range Transverse-Momentum Correlations and Radial Flow in Pb-Pb Collisions at the LHC.

Physical review letters·2026
Same author

Identification of a pathological feed-forward loop linking succinate, HIF1α signalling and vascular remodelling in abdominal aortic aneurysm.

European journal of clinical investigation·2026
Same author

Search for Quasiparticle Scattering in the Quark-Gluon Plasma with Jet Splittings in pp and Pb-Pb Collisions at sqrt[s_{NN}]=5.02  TeV.

Physical review letters·2025
Same author

First Measurement of A=4 Hypernuclei and Antihypernuclei at the LHC.

Physical review letters·2025
Same author

Targeting the pyruvate dehydrogenase kinase/pyruvate dehydrogenase axis modulates neutrophil and smooth muscle cell pathological responses and prevents abdominal aortic aneurysm formation.

Cardiovascular research·2025

A new fluorescence-based assay accurately quantifies HIV-protease activity using HPLC. This rapid and reproducible method enables sensitive detection of enzyme activity for HIV research.

Area of Science:

  • Biochemistry
  • Enzymology
  • Analytical Chemistry

Background:

  • Quantifying in vitro HIV-protease activity is crucial for drug development and understanding viral replication.
  • Existing methods may lack the sensitivity, speed, or reproducibility required for high-throughput screening or detailed kinetic studies.

Purpose of the Study:

  • To develop and validate a rapid, sensitive, and reproducible method for quantifying in vitro HIV-protease activity.
  • To utilize fluorescence detection coupled with High-Performance Liquid Chromatography (HPLC) for precise enzyme activity measurement.

Main Methods:

  • Developed a method based on the endoproteolytic conversion of N-dansyl-SQ-NYPIV (substrate) to N-dansyl-SQNY (product).
  • Employed N-dansyl group as a fluorescence label, assessing its minimal impact on kinetic parameters.

Related Experiment Videos

  • Utilized a single, rapid (3-minute) isocratic reverse-phase HPLC separation with fluorescence detection.

    • Main Results:

    • Achieved sensitive detection of as low as 0.2 pmol of product.
    • Demonstrated high reproducibility of the assay.
    • Confirmed that the N-dansyl label did not significantly alter peptide-enzyme kinetic parameters.

    Conclusions:

    • The developed HPLC-based fluorescence assay provides a rapid and sensitive method for quantifying in vitro HIV-protease activity.
    • The assay is highly reproducible and suitable for analyzing large sample numbers and conducting rigorous enzymological studies.
    • This method offers a valuable tool for HIV research, including drug discovery and mechanistic studies.