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

Equilibrium analysis of high affinity interactions using BIACORE

D G Myszka1, M D Jonsen, B J Graves

  • 1Huntsman Cancer Institute, University of Utah, 15 N. 2030 E., Room 2100, Salt Lake City, Utah, 84112-5330, USA. dmyszka@hci.utah.edu

Analytical Biochemistry
|January 12, 1999
PubMed
Summary
This summary is machine-generated.

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

Pit-1beta reduces transcription and CREB-binding protein recruitment in a DNA context-dependent manner.

The Journal of endocrinology·2005
Same author

The ABRF-MIRG'02 study: assembly state, thermodynamic, and kinetic analysis of an enzyme/inhibitor interaction.

Journal of biomolecular techniques : JBT·2004
Same author

Two of the five zinc fingers in the Zap1 transcription factor DNA binding domain dominate site-specific DNA binding.

Biochemistry·2003
Same author

Current and emerging commercial optical biosensors.

Journal of molecular recognition : JMR·2001
Same author

Survey of the year 2000 commercial optical biosensor literature.

Journal of molecular recognition : JMR·2001
Same author

Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding.

Cell·2001

This study presents a new method for BIACORE biosensors to measure high-affinity interactions. By adding analytes to the running buffer, researchers can collect equilibrium binding data for over 12 hours, improving kinetic analysis.

Area of Science:

  • Biophysical Chemistry
  • Biomolecular Interaction Analysis

Background:

  • BIACORE biosensors measure reaction kinetics and affinity constants for macromolecular interactions.
  • Standard injection methods limit association-phase data collection, especially for high-affinity interactions.

Purpose of the Study:

  • To develop a method to overcome limitations in BIACORE biosensor association-phase data collection.
  • To enable equilibrium analysis of high-affinity interactions.

Main Methods:

  • Utilized protein-DNA interactions as a model system.
  • Placed analyte directly into the running buffer for continuous supply to sensor surfaces (>12 hours).
  • Generated complete equilibrium binding profiles by altering analyte concentration and allowing reequilibration.

Related Experiment Videos

Main Results:

  • Demonstrated a simple and convenient method for BIACORE analysis.
  • Successfully collected association-phase data for extended periods.
  • Generated complete equilibrium binding profiles and confirmed reversibility of binding reactions.

Conclusions:

  • The described method overcomes limitations of standard BIACORE injection procedures for high-affinity interactions.
  • Enables direct measurement of equilibrium dissociation constants for very high affinity interactions.
  • Provides a robust approach for detailed kinetic and affinity analysis of biomolecular interactions.