Analyzing a kinetic titration series using affinity biosensors

Anal Biochem. 2006 Feb 1;349(1):136-47. doi: 10.1016/j.ab.2005.09.034. Epub 2005 Oct 13.


The classical method of measuring binding constants with affinity-based biosensors involves testing several analyte concentrations over the same ligand surface and regenerating the surface between binding cycles. Here we describe an alternative approach to collecting kinetic binding data, which we call "kinetic titration." This method involves sequentially injecting an analyte concentration series without any regeneration steps. Through a combination of simulation and experimentation, we show that this method can be as robust as the classical method of analysis. In addition, kinetic titrations can be more efficient than the conventional data collection method and allow us to fully characterize analyte binding to ligand surfaces that are difficult to regenerate.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Monoclonal / metabolism
  • Antibody Affinity*
  • Biosensing Techniques*
  • Kinetics
  • Phosphotransferases / metabolism
  • Prostate-Specific Antigen / immunology
  • Prostate-Specific Antigen / metabolism
  • Protein Binding
  • Software
  • Staurosporine / metabolism
  • Titrimetry*


  • Antibodies, Monoclonal
  • Phosphotransferases
  • Prostate-Specific Antigen
  • Staurosporine