Protein affinity chromatography substrate specificity

Affinity chromatography (AC) is a fractionation technique widely used in targeted proteomics or protein-protein interaction approaches. This method utilizes an interaction or affinity of a target protein to a substrate (or another protein) immobilized on a support matrix. Figure 17.4 demonstrates that AC can be incorporated into 3D proteomic approaches (Lee and Lee, 2004). Traditionally, AC has been used to purify, for example, carbohydratebinding proteins from Diplostomum pseudo-spathaceum (Mikes and Man, 2003) and the detoxification superfamily glutathione transferase (GST) from parasitic flatworms (Brophy and Barrett, 1990), specifically shown in... 

Affinity chromatography has become one of the standard techniques for the separation and purification of enzymes. It is based on the idea of utilizing the specific interaction between an enzyme and an immobilized substrate or inhibitor. The substrate is chemically bound to an inert support material, and when a sample is added to the column the enzyme binds to it to an extent depending on the strength of the interaction between the two. Other enzymes and proteins which do not interact with the substrate pass through the system with little or no retention. Removal of the enzyme from the stationary support is accomplished by changing the eluent so as to alter the enzyme-substrate interaction. The... 

Affinity Chromatography. Affinity chromatography is a very useful new technique that takes advantage of the natural affinity of a protein such as an enzyme for specific molecules, for instance, the substrate or a very effective inhibitor of that en-... 

Hjerten and co-workers prepared substrate-specific polyacrylamide gels by carrying out cross-linking polymerisation in the presence of proteins [47]. Polymerisation of acrylamide and 7V,A -methylenebisacrylamide was carried out in the presence of cytochrome C and haemoglobin as the templates. After removal of the templates using surfactants and acetic acid, substrate selectivities of the polymer gels were tested by chromatography. The imprinted polymers reportedly showed preferential affinities for their templates. 

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