Competitive enzyme-linked immunosorbent assay of the human cholesteryl ester transfer protein (CETP).
The present report describes the first competitive enzyme-linked immunosorbent assay (ELISA) for the cholesteryl ester transfer protein (CETP), an enzyme playing an important role in lipoprotein metabolism. This assay was developed with well-characterized TP1 anti-CETP monoclonal antibodies. The sensitivity of the ELISA assay was comparable with the sensitivity of the previously described radioimmunoassays since 1 ng of CETP per microwell of the immunoplate could be detected. Intra- and inter-assay coefficients of variation were 4% and 6%, respectively. This enzyme immunoassay provides a specific, sensitive and reproducible method for measuring CETP concentrations in various biological samples. Within normolipidemic subjects, the mean (+/- S.D.) of the plasma CETP concentration was 2.77 (+/- 0.59) micrograms/ml with a range of 1.87 to 4.23 micrograms/ml. When plasmas were supplemented with fatty acid-free albumin, the positive correlation observed between plasma CETP mass and CETP activity was improved, suggesting that plasma non-esterified fatty acids could play a role in modulating the activity of the cholesteryl ester transfer protein. When applied to the study of the binding of CETP to lipoprotein substrates, the enzyme immunoassay revealed that the experimental protocol used to separate lipoprotein fractions can have a great influence on the plasma distribution of CETP.
Guyard-Dangremont
V
V
Laboratoire de Biochimie des Lipoprotéines, INSERM CJF 93-10, Faculté de Médecine, Dijon, France.
Lagrost
L
L
Gambert
P
P
Lallemant
C
C
D000818:Animals; D000837:Anion Exchange Resins / Q000378:metabolism; D000911:Antibodies, Monoclonal; D001053:Apolipoproteins / Q000097:blood / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins / Q000097:blood; D006801:Humans; D008055:Lipids / Q000097:blood; D008297:Male; D051379:Mice; D012117:Resins, Synthetic; D012680:Sensitivity and Specificity; D014280:Triglycerides / Q000097:blood; D014461:Ultracentrifugation
D000837:Anion Exchange Resins; D000911:Antibodies, Monoclonal; D001053:Apolipoproteins; D001798:Blood Proteins; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D006023:Glycoproteins; D008055:Lipids; D012117:Resins, Synthetic; D014280:Triglycerides; C042855:Mono Q