Ultracentrifugation in swinging-bucket and fixed-angle rotors evaluated for isolation and determination of high-density lipoprotein subfractions HDL2 and HDL3.

摘要: We evaluated the density-gradient ultracentrifugation method in a swinging-bucket rotor (Anal Biochem 111, 149-157, 1981) slightly modified version for isolation and determination of high-density lipoprotein (HDL) subfractions. prestained serum with Coomassie Brilliant Blue R, which did not change hydrated densities lipoproteins, after only 2.2 X 10(8) gav . min obtained an equilibrium distribution lipoproteins along gradient. The density HDL 120 sera from apparently healthy persons patients different types hyperlipoproteinemia was bimodal. HDL2 could be isolated range 1.072-1.098 kg/L HDL3 at 1.100-1.176 kg/L, latter fraction being more heterogeneous. At solvent 1.100 we similar results HDL2-and HDL3-cholesterol by two fixed-angle rotors tube angles 15 degrees or 35 degrees. Independent technique, subfractionation d = resulted distinct stained entities than 1.125. In procedure, interference sinking pre-beta-lipoproteins minimized because, having between 1.058 1.075, they removed without aspirating HDL2. is both accurate precise. For HDL2- determined thawed frozen pool, CVs were 8.8 6.3%, respectively (n 18).