Kinetic studies of biological interactions by affinity chromatography.

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TitleKinetic studies of biological interactions by affinity chromatography.
Publication TypeJournal Article
Year of Publication2009
AuthorsSchiel, JE, Hage, DS
JournalJ Sep Sci
Volume32
Issue10
Pagination1507-22
Date Published2009 May
ISSN1615-9314
KeywordsChromatography, Affinity, Chromatography, High Pressure Liquid, Kinetics, Peptides, Proteins, Surface Plasmon Resonance, Time Factors
Abstract

The rates at which biological interactions occur can provide important information on the mechanism and behavior of such processes in living systems. This paper will discuss how affinity chromatography can be used as a tool to examine the kinetics of biological interactions. This approach, referred to here as biointeraction chromatography, uses a column with an immobilized binding agent to examine the association or dissociation of this agent with other compounds. The use of HPLC-based affinity columns in kinetic studies has received particular attention in recent years. Advantages of using HPLC with affinity chromatography for this purpose include the ability to reuse the same ligand within a column for a large number of experiments, and the good precision and accuracy of this approach. A number of techniques are available for kinetic studies through the use of affinity columns and biointeraction chromatography. These approaches include plate height measurements, peak profiling, peak fitting, split-peak measurements, and peak decay analysis. The general principles for each of these methods are discussed in this paper and some recent applications of these techniques are presented. The advantages and potential limitations of each approach are also considered.

DOI10.1002/jssc.200800685
Alternate JournalJ Sep Sci
PubMed ID19391173
PubMed Central IDPMC2782401
Grant ListR01 DK069629 / DK / NIDDK NIH HHS / United States
R01 DK069629 / DK / NIDDK NIH HHS / United States
R01 DK069629-03 / DK / NIDDK NIH HHS / United States
R01 GM044931 / GM / NIGMS NIH HHS / United States
R01 GM044931 / GM / NIGMS NIH HHS / United States
R01 GM044931-16 / GM / NIGMS NIH HHS / United States