Affinity elution

Chromatography of material such as proteins has occasionally been carried out in batch mode. Here the proteins are adsorbed onto a chromatographic medium by mixing the sample and the media in a common vessel prior to specific elution of the various adsorbed proteins. The mixture s components are subsequently eluted from the media by various types of elution schemes (e.g., salt and/or pH steps or gradients, specific affinity elution, etc.). 

Fifis, T. Scopes, R.K. Purification of 3-phosphoglycerate kinase from diverse sources by affinity elution chromatography. Biochem. J., 175, 311-319 (1978)... 

B 9. Describe how you would use affinity elution to remove the enzyme alcohol dehydrogenase bound to a Cibracron blue-agarose column. 

The bound RNA may then be eluted by unspecific denaturation with 7 M urea, 0.5 mM EDTA, 2% SDS, or heat. Another possibility is specific recovery of the bound RNA by affinity elution, in which the matrix is incubated with selection buffer containing free target molecules in excess relative to the immobilized target, so that target-binding RNA molecules are eluted. 

The affinity elution steps possible—for example, the peptide epitope when using an antibody, imidazole or EDTA when using a His-tag, or specific sugar residue when using a lectin column to purify glycoproteins—are likely to improve the specificity of the isolation process. 

Elution of the antibodies with a linear concentration gradient of the hapten, fractionated the antibodies according to their affinity. Elution in this manner permits the study of antibody populations selected on the basis of their affinity. 

Cross-reacting antigens (or antibodies) with lower affinities, elute more rapidly through repeated washings than the specific antigen (or antibody). Successive washings can, therefore, be beneficial for the elimination of cross-reacting immunoreactants. 

In our laboratory, we feel that the scientific and practical use of these inhibitors, by no means, has reached a peak. An obvious need is to try different ligands for affinity elution. Therefore, a new class of transition state analogs was synthesized and tested for their inhibitory potential against JHE, cholinesterase and malathionase. 

P, P -Di(adenosine-5 )pentaphosphate has been used for the specific affinity elution of adenylate kinase from a number of proteins bound unspecifically to blue-Sepharose in a simple, rapid purification. ... 

Affinity elution chromatography, using 3-phosphoglycerate as eluant and carboxymethylcellulose as support matrix, has been applied to the purification of 3-phosphoglycerate kinase from a variety of sources. ... 

Investigation of suitability of the matrix for enzyme immobilization Isolation of the enzymes by specific affinity elution Active immobilized enzyme study of 518... 

The flow rate used is an important factor in affinity elution. The lowest flow rate acceptable to the chromatography conditions is used to obtain maximum recovery and resolution with a minimum dilution of the sample. This is particularly important if several proteins have bound to the affinity adsorbent. The direction of the flow... 

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