Kinetic studies of biological interactions by affinity chromatography.
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.
Schiel
John E
JE
Department of Chemistry, University of Nebraska, Lincoln, NE 68588-0304, USA.
Hage
David S
DS
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D007700:Kinetics; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D013997:Time Factors
D010455:Peptides; D011506:Proteins