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Isolation and quantitation of apolipoproteins A-I and A-II from human high-density lipoproteins by fast-protein liquid chromatography》.pdf
Clinica Chimica Acta, 169 (1987) 249-254 249 Elsevier CCA 03974 Isolation and quantitation of apolipoproteins A-I and A-II from human high-density lipoproteins by fast-protein liquid chromatography Peter Weisweiler with the technical assistance of Christine Fried1 and Margrit Ungar iphar-Institute for Clinical Pharmacology, Munich (FRG) (Received 31 March 1987; revision received 30 May 1987; accepted after revision 3 August 1987) Key words: Apolipoproteins A-I and A-II; Fast-protein liquid chromatography Summary Apolipoproteins (APO) A-I and A-II from human high-density lipoproteins (HDL) were isolated and quantified by fast-protein liquid chromatography using a Superose 12 column (gel filtration) followed by a Mono Q column (anion-ex- changer). The separation times were 45 rnin and 15 min, respectively. Identities of both apolipoproteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by double immunodiffusion. Peak areas after Mono Q chro- matography increased linearly with concentration for samples from 0.5 to 10 mg of protein with a mean ratio of Apo A-I to Apo A-II of 3.46 f 0.71 in normolipopro- teinemic subjects. This precise technique is an alternative for preparing and quanti- fying both HDL apolipoproteins. Introduction The procedures which are in common use to isola
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