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From gel filtration to biosensor technology: the development of chromatography for the characterization of protein interactions
Author(s) -
Winzor Donald J.
Publication year - 2000
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/1099-1352(200009/10)13:5<279::aid-jmr506>3.0.co;2-k
Subject(s) - chemistry , size exclusion chromatography , partition equilibrium , affinity chromatography , chromatography , affinities , biosensor , equilibrium constant , characterization (materials science) , filtration (mathematics) , ionic strength , ion chromatography , organic chemistry , nanotechnology , materials science , aqueous solution , stereochemistry , biochemistry , statistics , mathematics , enzyme
The objective of this review is to summarize the development of chromatographic techniques for the determination of reaction stoichiometries and equilibrium constants for solute interactions of biological importance. Gel chromatography is shown to offer a convenient means of characterizing solute self‐association as well as solute–ligand interactions. Affinity chromatography is an even more versatile method of characterizing interactions between dissimilar reactants because the biospecificity incorporated into the design of the affinity matrix ensures applicability of the method regardless of the relative sizes of the two reactants. Adoption of different experimental strategies such as column chromatography, simple partition equilibrium experiments and biosensor technology has created a situation wherein affinity chromatography affords a means of characterizing the whole range of reaction affinities—from relatively weak interactions (binding constants less that 10 3 M −1 ) to tight interactions with binding constants greater than 10 9 M −1 . In addition to its established prowess as a means of solute separation and purification, chromatography thus also possesses considerable potential for investigation of the functional roles of the purified reactants—an endeavour that requires characterization as well as identification of the interactions responsible for a physiological phenomenon. Copyright © 2000 John Wiley & Sons, Ltd.