Abstract
The rapid separation of seven urea-soluble apolipoprotein species from delipidated human serum very low density lipoproteins (VLDL) and high density lipoproteins (HDL) has been achieved by high-performance liquid chromatography on an anion-exchange column of Syn-Chropak AX 300. Effluent chromatographic peaks were detected by absorbance at 280 nm in a flow-through cell. Peaks corresponding to apolipoproteins AI 1, AI 2, AII, CI, CII, CIII 1, and CIII 2 were identified by amino acid analysis, gel electrophoresis, and isoelectric focusing. Maximum efficient loading of semipreparative columns (250 × 9.0 mm) was established to be ca. 20 mg HDL apolipoprotein. Minimum detectable protein was shown to be ca. 1 μg on an analytical-scale column (300 × 4.5 mm). Chromatographic resolution is comparable to that of conventional DEAE-cellulose column chromatography. The ratio of apoAI 1 to apoAI 2 was considerably greater in high-performance liquid chromatography, suggesting that the variants seen in conventional chromatography and isoelectric focusing are in part artifactual.
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