[Isolation of isoforms of apolipoprotein CIII from human serum by chromatofocusing].
This study aimed to isolate isoforms of apolipoprotein (apo) C III from human serum. 24-hour fasting serum from normal and hyperlipidemic subjects was pooled and subjected to ultracentrifugation at plasma density for 20 hours. Very low density lipoprotein (VLDL) was collected at density of d < 1.006 g/ml, and it was delipidated by ethanol and ether. The delipidated apo-VLDL was dissolved in a solution containing 7.2 mol/L urea and 20 mmol/L dithiothreitol. The insoluble apo B was removed by centrifugation. The soluble apo-VLDL was applied to PBE94 column, and eluted with elution buffer containing polybuffer 74 and 8 mol/L urea (1:8, pH4.0). After pooled, the eluted peaks of apolipoproteins were applied to column chromatography of hydroxylapatite to remove the polybuffer. The purified isoforms of apoC III and the purified apo C I, C II and E, were characterized by isoelectrofocusing and west blot. The results showed that the purified apoC III1, C III2, and C II were pure.
Department of Biochemistry, School of Basic Medical Sciences, WCUMS, Chengdu 610041.
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D053305:Apolipoprotein C-III; D001056:Apolipoproteins C / Q000097:blood* / Q000145:classification / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D006801:Humans
D053305:Apolipoprotein C-III; D001056:Apolipoproteins C