A simple method for assessing copper-mediated oxidation of very-low-density lipoprotein isolated by rapid ultracentrifugation.
The association of very-low-density lipoprotein (VLDL) with atherosclerosis remains controversial. However, studies have shown that oxidative modification of VLDL can promote foam cell formation, leading to the development of atherosclerosis. A rapid method is described which will allow the significance of VLDL oxidation to be assessed in clinical studies. VLDL was isolated from heparinized plasma by a 1-h, single spin ultracentrifugation. Total protein was standardized to 25 mg/L. Oxidation was promoted by the addition of copper ions (17.5 mumol/L, final concentration) incubated at 37 degrees C. Conjugated diene production was followed at 234 nm. Total assay preparation time was 2 h. Urate greatly inhibited the oxidation of VLDL and was successfully removed by size exclusion chromatography. VLDL isolated from frozen plasma (-70 degrees C) was stable for 15 weeks. This simple, rapid method for the isolation of VLDL may be applied to assess the significance of VLDL oxidation in disease.
Department of Clinical Biochemistry, Queen's University, Belfast, UK.
J
J
McEneny
E R
ER
Trimble
I S
IS
Young
D003300:Copper / Q000378:metabolism*; D006801:Humans; D008054:Lipid Peroxides / Q000032:analysis; D008079:Lipoproteins, VLDL / Q000302:isolation & purification* / Q000378:metabolism; D010084:Oxidation-Reduction; D012016:Reference Values; D013997:Time Factors; D014461:Ultracentrifugation / Q000379:methods*
D008054:Lipid Peroxides; D008079:Lipoproteins, VLDL; D003300:Copper