Determination of binding constants by affinity chromatography.
This review summarizes developments in the use of affinity chromatography to characterize biospecific interactions in terms of reaction stoichiometry and equilibrium constant. In that regard, the biospecificity incorporated into the design of the experiment ensures applicability of the method regardless of the sizes of the reacting solutes. By the adoption of different experimental strategies (column chromatography, simple partition equilibrium, solid-phase immunoassay and biosensor technology protocols) quantitatiative affinity chromatography can be used to characterize interactions governed by an extremely broad range of binding affinities. In addition, the link between ligand-binding studies and quantitative affinity chromatography is illustrated by means of partition equilibrium studies of glycolytic enzyme interactions with muscle myofibrils, an exercise which emphasizes that the same theoretical expressions apply to naturally occurring examples of affinity chromatography in the cellular environment.
Winzor
Donald J
DJ
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Qld 4072, Australia. d.winzor@mailbox.uq.edu.au
D015374:Biosensing Techniques; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000378:metabolism; D006019:Glycolysis; D008433:Mathematics; D009210:Myofibrils / Q000737:chemistry / Q000378:metabolism; D011863:Radioimmunoassay; D012996:Solutions
D012996:Solutions; D005634:Fructose-Bisphosphate Aldolase