+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Development of a homogeneous assay to measure remnant lipoprotein cholesterol

Clinical Chemistry 53(12): 2128-2135

Development of a homogeneous assay to measure remnant lipoprotein cholesterol

Background: Quantification of triglyceride-rich lipoprotein (TRL) remnants is useful for risk assessment of coronary artery disease and the diagnosis of type III hyperlipoproteinemia. Although an immunoseparation procedure for remnant-like particle cholesterol has been evaluated extensively in recent years, available methods for measuring TRL remnants have not achieved wide use in routine laboratory practice, suggesting a need for a homogeneous assay that can measure TRL remnant cholesterol in serum or plasma without pretreatment.Methods: We screened for suitable surfactants that exhibited favorable selectivity toward the VLDL remnant (VLDLR) fraction, including intermediate-density lipoproteins (IDLs). We investigated the principal characteristics of this assay by gel filtration of lipoproteins and their particle size distribution. We developed a simple assay and evaluated its performance with the Hitachi-7170 analyzer.Results: Polyoxyethylene-polyoxybutylene block copolymer (POE-POB) exhibited favorable selectivity toward VLDLR and IDL fractions. POE-POB removed apolipoprotein (apo) E and apo C-III from IDL particles in the presence of cholesterol esterase (CHER), and the particle size distribution of IDLs became smaller after the reaction. These results revealed that IDL particles are specifically modified in the presence of CHER and POE-POB, making their component cholesterol available for enzymatic assay. Addition of phospholipase D improved the reactivity toward chylomicron remnants (CMRs). We found a high correlation [y = 1.018x- 0.01 mmol/L, r = 0.962 (n = 160)] between the proposed assay and the immunoseparation assay in serum from healthy individuals.Conclusion: The homogeneous assay described in this report can measure TRL remnant cholesterol, including CMRs, VLDLRs, and, IDLs, with high sensitivity and specificity. (C) 2007 American Association for Clinical Chemistry.

Accession: 020840610

PMID: 17901111

DOI: 10.1373/clinchem.2007.092296

Related references

Murano, T.; Kiuchi, M.; Moteki, M.; Miyashita, Y.; Watanabe, H.; Hiruta, N.; Shirai, K., 2008: Remnant lipoprotein cholesterol level measured by homogeneous assay reflects the quantity of intermediate-density lipoprotein. Remnant lipoprotein can be measured by immunoseparation assay and polyacrylamide gel disc electrophoresis; however, these methods have some problems, such as a lack of specificity and being technically complicated. Recently, a new method, the remn...

Nakada, Y.; Kurosawa, H.; Tohyama, J-Ichiro.; Inoue, Y.; Ikewaki, K., 2007: Increased remnant lipoprotein in patients with coronary artery disease--evaluation utilizing a newly developed remnant assay, remnant lipoproteins cholesterol homogenous assay (RemL-C). Remnant lipoprotein is an emerging risk factor for coronary artery disease (CAD); however, the development of a specific remnant lipoprotein assay has struggled due to its heterogeneous nature. This study aimed to evaluate the clinical importance...

Nagashima, M.; Mori, Y.; Morita, R.; Takada, M.; Lee, S.; Taira, T.; Miyauchi, K.; Kuwata, H.; Kayahara, N.; Hirano, T., 2009: Remnant lipoprotein cholesterol homogenous assay (RemL-C) is closely associated with very-low-density lipoprotein remnants: comparison with the immunoseparation assay. Remnant-like particle-cholesterol (RLP-C) is recognized as a risk factor for cardiovascular disease. As an alternative to the immunoseparation assay widely used for the measurement of RLP-C, a new remnant lipoprotein-C homogenous assay (RemL-C) is...

Ito, Y.; Satoh, N.; Ishii, T.; Kumakura, J.; Hirano, T., 2015: Development of a homogeneous assay for measurement of high-density lipoprotein-subclass cholesterol. Several studies have suggested that measurement of high-density lipoprotein (HDL) 2 and HDL3 subfractions might be more useful for evaluating coronary risk than total HDL-cholesterol (C). However, methods of measuring HDL2 and HDL3 are quite labor...

Kishi, K.; Ochiai, K.; Ohta, Y.; Uemura, Y.; Kanatani, K.; Nakajima, K.; Nakamura, M., 2002: Highly sensitive cholesterol assay with enzymatic cycling applied to measurement of remnant lipoprotein-cholesterol in serum. Background: Remnant lipoprotein-cholesterol (RLP-C) concentrations in sera of healthy individuals are very low (0.080-0.437 mmol/L), making conventional cholesterol methods poorly suited to this purpose. We have developed a highly sensitive choles...

Herzum, I.; Giehl, C.; Soufi, M.; Junclas, H.; Wahl, H-Guenther., 2007: Interference in a homogeneous assay for low-density lipoprotein cholesterol by lipoprotein X. Background: Homogeneous assays for cholesterol in low-density lipoprotein (LDL) are currently in wide use for guideline-based diagnosis and monitoring of dyslipaemic or coronary conditions. In some sera from patients with impaired liver function,...

Devaraj, S.; Vega,; Scott, G.M.; Ishwarlal, J., 1996: Evaluation of a rapid remnant lipoprotein cholesterol assay in atherosclerosis. Circulation 94(8 SUPPL ): I121

Schaefer, E.J., 2009: Limitations of automated remnant lipoprotein cholesterol assay for diagnostic use. Clinical Chemistry 55(11): 2061-2; Author Reply 2062-3

Devaraj, S.; Vega, G.L.; Grundy, S.M.; Jialal, I., 1997: Evaluation of a new assay for measurement of remnant lipoprotein cholesterol in patients with coronary artery disease. Clinical Chemistry 43(6 PART 2): S266

Sakurabayashi, I.; Daimon, M.; Kamiyama, K.; Takanashi, K.; Suzuki, Y.; Watanabe, M.; Kubono, K.; Nakajima, K., 2003: Screening for Type III and other familial hyperlipidemia among Japanese hyperlipidemic subjects by using the remnant lipoprotein cholesterol assay. Clinical Chemistry 49(S6): A163, June