A new method for hdl particle sizing by polyacrylamide...

C - Chemistry – Metallurgy – 07 – K

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C07K 1/14 (2006.01) C07K 14/435 (2006.01) G01N 1/34 (2006.01) G01N 1/42 (2006.01) G01N 27/447 (2006.01) G01N 33/52 (2006.01) G01N 33/92 (2006.01) G01N 15/02 (2006.01)

Patent

CA 2337547

Low plasma levels of high-density lipoprotein (HDL) cholesterol have been associated with an increased risk of coronary heart disease (CHD). HDL are very heterogeneous with respect to size and apolipoprotein content and the objective of the present study was to develop a method to generate lipid-stainable standards that would allow for the assessment of HDL particle size from whole plasma using polyacrylamide gradient gel electrophoresis (PAGGE). Lipid-stainable plasma standards were obtained by subjecting isolated red blood cells to hemolysis by either freezing at - 20°C or -80°C overnight or to rapid exposure to liquid nitrogen and mixing the hemolysis products with plasma aliquots. All three methods were highly reproducible in producing Sudan black lipid-stainable standards ranging from 75 .ANG. to 200 .ANG.. With the use of these standard bands, HDL particle size was assessed with intra and inter assay coefficients of variation below 0.8% and 1.2% respectively. This use of lipid-stainable HDL standards simplifies the assessment of HDL particle size by PAGGE using whole plasma, without having to go through cost and time consuming; ultracentrifugation procedures.

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