Prog. Lipid Res. Vol. 30, No. 2/3, pp. 181-187, 1991 0163-7827/91/$0.00 + 0.50 Printed in Great Britain. All rightsreserved © 1991 Pergamon Press plc SEPARATION OF ApoA- AND ApoB-CONTAINING LIPOPROTEINS OF HUMAN PLASMA BY AFFINITY CHROMATOGRAPHY ON CONCANAVALIN A MARCELO TAVELLA,*~f PETAR ALAUPOVIC,~ CAROLYN KNIGHT-GIBSON,~ HORACIO TOURNIER,~" GUILLERMO SCHINELLAt and OSVALDO MERCURIJf t lnstituto de InvestigacionesBioquimicas de La Plata, School of Medicine, 1900 La Plata, Argentina ~Lipoprotein and Atherosclerosis Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, U.S.A. CONTENTS ABBREVIATIONS 181 I. INTRODUCTION 181 II. SEPARATION OF APOA- ANDAPoB-CONTAIN1NG LIPOPROTEINS OF HUMAN PLASMA BY AFFINITY CHROMATOGRAPHY ON CONA 183 A. Column chromatography on ConA 183 1. Description of subjects and patients 183 2. Column chromatography 184 B. Characterization of APOA- and ApoB-containing lipoproteins 184 1. Lipids and apolipoproteins 184 2. Fatty acid composition 185 IlL CONCLUSION 186 ACKNOWLEDGEMENTS 186 REFERENCES 186 ABBREVIATIONS VLDL---very low density lipoproteins LDL--low density lipoproteins HDL---high density lipoproteins Apo--apolipoprotein LP--lipoprotein ConA----concanavalin A I. INTRODUCTION The chemical, metabolic and kinetic heterogeneity of operationally defined plasma liPoprotein classes ~'6'34'39'44'49 and the recognition that this heterogeneity is mainly due to the presence of discrete lipoprotein families characterized by distinct apolipoprotein composition but similar physical properties, 1'2'n'21'31 necessitated the introduction of an alternative system for classifying plasma lipoproteins.~'2 This classification system views plasma lipoproteins as a mixture of discrete lipoprotein families, each of which represents a polydisperse system of particles heterogeneous with respect to their physical properties (size, density and charge distribution) but homogeneous with respect to their specific qualitative apolipoprotein composition. Studies on the quantification of apolipoproteins have shown that ApoA (ApoA-I + ApoA-II) and ApoB are the major apolipoproteins distributed monomodaly along the density spectrum from 0.92 to 1.21 g/ml. 2's'n Whereas, ApoA occurs mainly in high density lipoprotein (HDL) particles, ApoB is present in lipoprotein particles of very low (VLDL) and low densities (LDL). 2,3 The fractionation of ApoA- and ApoB-containing lipoproteins by sequential immunoprecipitation or sequential immunoaffinity chromatography has disclosed that the former lipoproteins consist of three distinct lipoprotein families, LP-A-I, LP-A-I: A-II and LP-A-II, and the latter ones of five major lipoprotein families, LP-B, LP-B: C, LP-B: C: E, LP-B: E and LP-A-II: B: C: D: E.2'4"sa°,~6 On the basis of these findings, plasma lipoproteins are classified *Corresponding address: Mareelo Tavella, M.D., Instituto de Investigaciones Bioquimieas, La Plata (INIBIOLP), (1900) La Plata, Argentina. 181