A new method for HDL particle sizing by polyacrylamide gradient gel electrophoresis using whole plasma

Low plasma levels of HDL cholesterol have been associated with an increased risk of coronary heart disease. HDL particles are heterogeneous with respe ct to size and apolipoprotein content. The objective of the present study w as to develop a method to generate lipid-stainable calibrators that would a llow the assessment of HDL particle size from whole plasma, using polyacryl amide gradient gel electrophoresis (PAGGE). Lipid-stainable HDL calibrators were obtained by subjecting isolated red blood cells to hemolysis either b y freezing at - 20 or - 80 degreesC overnight or by rapid exposure to liqui d nitrogen and mixing of the hemolysis products with plasma aliquots. All t hree methods were highly reproducible in producing Sudan black lipid-staina ble HDL calibrators ranging from 75 to 200 A. The assessment of HDL particl e size with these lipid-stainable HDL calibrators was also highly reproduci ble, with a coefficient of variation below 5.5%.ie These lipid-stainable HD L calibrators simplify the assessment of HDL particle size by PAGGE using w hole plasma, without the need for costly, time-consuming ultracentrifugatio n procedures.