Evaluation of an immunoseparation method for quantitative measurement of remnant-like particle-cholesterol in serum and plasma

Substantial evidence indicates that triglyceride-rich lipoprotein remnants are atherogenic. Additional research has, however, been limited by available methods for separation and quantification of remnants. We have evaluated an immunoseparation assay developed to measure cholesterol in remnant-like...

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Published in:Clinical chemistry (Baltimore, Md.) Md.), 1998-12, Vol.44 (12), p.2490-2498
Main Authors: Leary, Elizabeth Teng, Wang, Tao, Baker, Daniel J, Cilla, Donald D, Zhong, Jianhua, Warnick, G. Russell, Nakajima, Katsuyuki, Havel, Richard J
Format: Article
Language:English
Subjects:
Adolescent
Adult
Aged
Apolipoproteins - blood
Biological and medical sciences
Blood Specimen Collection
Cardiac dysrhythmias
Cardiology. Vascular system
Cholesterol
Coronary Angiography
Coronary Disease - blood
Coronary Disease - diagnostic imaging
Female
Heart
Humans
Lipoproteins - blood
Male
Medical sciences
Middle Aged
Reagent Kits, Diagnostic
Reference Values
Triglycerides - blood
United States
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Summary:Substantial evidence indicates that triglyceride-rich lipoprotein remnants are atherogenic. Additional research has, however, been limited by available methods for separation and quantification of remnants. We have evaluated an immunoseparation assay developed to measure cholesterol in remnant-like particles (RLP-C). This method uses monoclonal antibodies to human apolipoproteins B-100 and A-I to remove most of the apolipoprotein B-100-containing lipoproteins (namely LDL and nascent VLDL) and apolipoprotein A-I-containing lipoproteins (namely chylomicrons and HDL), leaving behind a fraction of triglyceride-rich lipoproteins, including chylomicron and VLDL remnants, both of which are enriched in apolipoprotein E. Cholesterol in the unbound fraction is measured with a sensitive enzymatic assay. The RLP-C concentration was highly correlated with total triglyceride-rich lipoproteins (sum of VLDL-cholesterol and IDL-cholesterol) separated by ultracentrifugation and by polyacrylamide gel electrophoresis (r = 0.86 and 0.76, respectively). The within-run and run-to-run imprecision (CV) of the assay was approximately 6% and 10%, respectively. The assay was not affected by hemoglobin up to 5000 mg/L (500 mg/dL), bilirubin up to 342 mmol/L (20 mg/dL), glucose up to 67 mmol/L (1200 mg/dL), or ascorbic acid up to 170 mmol/L (3.0 mg/dL). In 726 subjects (men, n = 364; women, n = 362) in the US, the 75th percentiles of RLP-C concentration were 0.17 mmol/L (6.6 mg/dL) and 0.23 mmol/L (8.8 mg/dL) in sera obtained after overnight fasting or randomly, respectively. A group of 151 patients from nine US centers and one Canadian center with coronary artery atherosclerosis established by angiography had higher median RLP-C concentrations than 302 gender- and age-matched controls (P
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/44.12.2490