Semi-automated rapid isoelectric focusing of apolipoproteins C from human plasma using Phastsystem and immunofixation

Clin Chem Lab Med. 1999 Jun;37(6):643-8. doi: 10.1515/CCLM.1999.100.

Abstract

Apolipoproteins (apo) C-I, C-II, and C-III play crucial roles in intravascular lipid metabolism. Whereas apo C-II is an obligate cofactor for lipoprotein lipase, apo C-III was shown to inhibit its action. Apo C-I can be a potent cofactor of human lecithin:cholesterol acyltransferase. Structural mutants and deficiencies of apo C-II lead to hypertriglyceridemia. A similar phenotype is associated with apo C-III mutants and is inducible by overexpression of human apo C-III in transgenic animals. No structural variant has so far been reported for apo C-I. The present paper describes a rapid semi-automated procedure for isoelectric focusing analysis of these C-apolipoproteins from whole plasma or serum and their visualization by immunofixation and silver staining. The procedure allows detection of charged variants of C-apolipoproteins. As applied to 295 patients with coronary heart disease and 85 controls, it also serves to detect deficiency syndromes of these apolipoproteins. The procedure provides reliable, easy and quick analysis of C-apolipoproteins applicable as a routine or screening procedure not restricted to specialized laboratories.

MeSH terms

  • Acrylic Resins
  • Apolipoproteins C / blood*
  • Apolipoproteins C / isolation & purification
  • Autoanalysis
  • Blood Protein Electrophoresis / methods
  • Coronary Disease / blood
  • Humans
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing / methods
  • Precipitin Tests
  • Reproducibility of Results

Substances

  • Acrylic Resins
  • Apolipoproteins C
  • polyacrylamide gels