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Isoelectric focusing of apolipoproteins in immobilized pH gradients: Improved determination of apolipoprotein Ephenotypes

✍ Scribed by Dr. Manfred W. Baumstrak; Aloys Berg; Martin Halle; Joseph Keul

Publisher
John Wiley and Sons
Year
1988
Tongue
English
Weight
523 KB
Volume
9
Category
Article
ISSN
0173-0835

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✦ Synopsis

A method for isoelectric focusing of apolipoprotein E in an immobilized pH gradient with added carrier ampholytes has been developed. This method is an improvement over conventional isoelectric focusing of apolipoprotein E with respect to resolution, reproducibility, and simplicity. Since monosialo isoforms are resolved from the normally cofocusing asialo isoforms, unique patterns are obtained for all 6 common apolipoprotein E phenotypes. The method can also be applied to the screening of apolipoprotein A and C isoforms. Delipidated very low density lipoproteins (VLDL) have been used as the source of apolipoprotein E and C. Apolipoprotein A isoforms were focused directly from detergent-treated serum. Immunodetection of apolipoprotein E using capillary transfer was found to be compatible with the described method.

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Apolipoproteins C are involved in many ways in the metabolism of plasma lipoproteins. Apolipoproteins C from the delipidated VLDL of 35 controls and 165 normo- and hyperlipoproteinemic patients were analyzed by isoelectric focusing on an immobilized pH gradient, pH 4.0-5.0, with 7 M urea, which rais