Discovery and Development of Hydrophobic Interaction Chromatography

The chromatographic puriRcation of proteins on spe-ciRcally synthesized hydrophobic solid supports was first reported independently by Yon and Shaltiel in 1972. In both cases the hydrophobic matrix consisted of agarose to which aminoalkane derivatives have been coupled by the CNBr method. Yon synthesized mixed hydrophobic-charged gels (aminodecyl-, or N-3-carboxypropionyl)aminodecyl-agarose) with an alkyl residue to charge ratio of at least 1 : 1 for the adsorption of lipophilic proteins such as bovine serum albumin or aspartate transcarbamoylase. These proteins were adsorbed at low ionic strength at the isoelectric point and eluted at acidic or alkaline pH by charge repulsion. The surprising result in Shal-tiel's experiments was that a very normal hydrophilic enzyme, phosphorylase b, could be puriRed on hydrocarbon-coated agaroses to near homogeneity in one step, implicitly questioning the general doctrine of the time that all hydrophobic amino acids are buried in the interior of proteins. Phosphorylase was adsorbed at low ionic strength on immobilized butyl residues which had no resemblance to the substrates of the enzyme (excluding affinity chromatography) and was eluted by a 'deforming buffer' which imposed a limited conformational change on the enzyme. Taken together with Shaltiel's systematic approach of grading the hydrophobicity of the gels via an immobilized homologous hydrocarbon series, the immediate

Solar Panel Basics

Solar Panel Basics

Global warming is a huge problem which will significantly affect every country in the world. Many people all over the world are trying to do whatever they can to help combat the effects of global warming. One of the ways that people can fight global warming is to reduce their dependence on non-renewable energy sources like oil and petroleum based products.

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