Determination of immuno-reactive rabbit apolipoprotein B-48 in serum by ELISA.
Apolipoprotein B-48 (apoB-48) is produced by the small intestine, is associated with chylomicrons, and appears to be a suitable marker for clinical studies of postprandial dynamics of lipoproteins. It is also associated with cardiovascular risk factors. We have developed an assay system to quantify immuno-reactive apoB-48 in rabbit serum or plasma. A microtiter-plate was coated with monoclonal antibody raised against human apoB-48 C-terminal specific decapeptide that has high homology to the rabbit C-terminal sequence. Appropriate ELISA standard curves were obtained using apoB-48 extracted from rabbit serum by immuno-affinity chromatography. No cross-reactivity was found with apoB-100 in western blot analyses. Intra- and inter-assay CVs were less than 3%. Recovery of rabbit apoB-48 spiked in serum was within 93.4-105%. ApoB-48 levels in healthy controls rabbits fed a normal diet were within the range of 0.903-1.09 microg/ml (mean +/- SD: 1.03 +/- 0.084 microg/ml). In healthy animals, the blood apoB-48 level was markedly increased by a high fat diet and in the postprandial state in parallel with serum triglyceride. Ezetimibe, cholesterol absorption inhibitor, given orally to rabbits fed on a high fat diet blocked further increase of blood levels of apoB-48 and triglyceride. This method for measuring apoB-48 using the monoclonal antibody is simple, reliable, and suitable for routine analyses.
Kinoshita
Makoto
M
Department of Internal Medicine, Teikyo University School of Medicine, Itabashi, Tokyo, Japan.
Matsushima
Teruhiko
T
Mashimo
Yamato
Y
Kojima
Masaaki
M
Kigure
Megumi
M
Teramoto
Tamio
T
D000284:Administration, Oral; D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D053283:Apolipoprotein B-48 / Q000037:antagonists & inhibitors / Q000097:blood* / Q000276:immunology; D001384:Azetidines / Q000494:pharmacology; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000069438:Ezetimibe; D008297:Male; D011817:Rabbits; D012016:Reference Values; D015203:Reproducibility of Results; D014280:Triglycerides / Q000037:antagonists & inhibitors / Q000097:blood
D000924:Anticholesteremic Agents; D053283:Apolipoprotein B-48; D001384:Azetidines; D014280:Triglycerides; D000069438:Ezetimibe