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.
McEneny
J
J
Department of Clinical Biochemistry, Queen's University, Belfast, UK.
Trimble
E R
ER
Young
I S
IS
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