In vivo Pharmacology Group, Oligonucleotide Therapeutics Unit, Pfizer Inc., Cambridge, MA.
In vivo Pharmacology Group, Oligonucleotide Therapeutics Unit, Wake Forest University School of Medicine, Winston-Salem, NC.
J Lipid Res. 2011 Jun;52(6):1265-1271. doi: 10.1194/jlr.D011148. Epub 2011 Mar 2.
Apolipoprotein CIII (apoCIII), a major constituent of triglyceride-rich lipoprotein, has been proposed as a key contributor to hypertriglyceridemia on the basis of its inhibitory effects on lipoprotein lipase. Many immunochemical methods have been developed for human apoCIII quantification, including ELISA. However, a sensitive and quantitative assay for nonhuman primates is not commercially available. We developed a sensitive, quantitative, and highly specific sandwich ELISA to measure apoCIII in both nonhuman primate and human serum. Our assay generates a linear calibration curve from 0.01 μg/ml to 10 μg/ml using an apoCIII standard that was purified from cynomolgus monkey serum. It is highly reproducible (intra- and interplate CV < 5% and < 8%, respectively), sensitive enough to distinguish 10% difference of apoCIII present in serum, and has no interference from purified human apolipoprotein AI, AII, B, CI, CII, or E. The same assay can also be used to measure human apoCIII with a linear calibration curve from 0.005 μg/ml to 1 μg/ml using purified human apoCIII as the standard. This fast and highly sensitive ELISA could be a useful tool to investigate the role of apoCIII in lipoprotein transport and cardiovascular disease.
载脂蛋白 CIII（apoCIII）是富含甘油三酯的脂蛋白的主要成分，基于其对脂蛋白脂肪酶的抑制作用，被认为是导致高甘油三酯血症的关键因素。已经开发出许多用于人 apoCIII 定量的免疫化学方法，包括 ELISA。然而，商业化的非人灵长类动物敏感和定量检测方法尚未问世。我们开发了一种灵敏、定量和高度特异的夹心 ELISA 来测量非人类灵长类动物和人血清中的 apoCIII。我们的测定方法使用从食蟹猴血清中纯化的 apoCIII 标准品，在 0.01 μg/ml 至 10 μg/ml 的范围内生成线性校准曲线。它具有高度可重复性（内板和间板 CV 分别<5%和<8%），足以区分血清中存在的 apoCIII 的 10%差异，并且不受纯化的人载脂蛋白 AI、AII、B、CI、CII 或 E 的干扰。同样的测定方法也可以用于测量人 apoCIII，使用纯化的人 apoCIII 作为标准品，在 0.005 μg/ml 至 1 μg/ml 的范围内生成线性校准曲线。这种快速且高度敏感的 ELISA 可能是研究 apoCIII 在脂蛋白转运和心血管疾病中作用的有用工具。
