Separation of the apoprotein components of human very low density lipoproteins by ion-paired, reversed-phase high performance liquid chromatography

W. S. Hancock, C. A. Bishop, Antonio Gotto, D. R.K. Harding, S. M. Lamplugh, J. T. Sparrow

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

A number of crude apolipoprotein samples isolated from human very low density lipoproteins (VLDL) were analyzed by reversed phase high performance liquid chromatography. The mobile phase consisted of a 1% solution of the polar ion-pairing reagent triethylammonium phosphate. A slow, nonlinear gradient of acetonitrile (37-42%) was used to elute the apolipoproteins. The order of elution was as follows: apolipoprotein Cx, apolipoprotein C-I, apolipoprotein C-III2, apolipoprotein C-III1, apolipoprotein C-III0 and apolipoprotein C-II. This order is consistent with the known polarity of the proteins, i.e., the most nonpolar, apolipoprotein C-II, was the last to be eluted, whereas apolipoprotein C-I, with the lowest nonpolar surface area eluted first. The recovery of the individual apolipoproteins was 80-95% and the individual peaks were characterized by amino acid analysis, UV absorption spectra and chromatography of pure protein standards.

Original languageEnglish (US)
Pages (from-to)250-259
Number of pages10
JournalLipids
Volume16
Issue number4
DOIs
StatePublished - Apr 1 1981

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Cell Biology

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