Kinoshita Makoto, Kojima Masaaki, Matsushima Teruhiko, Teramoto Tamio
Department of Internal Medicine, Teikyo University School of Medicine, Itabashi, Tokyo 173-8605 Japan.
Clin Chim Acta. 2005 Jan;351(1-2):115-20. doi: 10.1016/j.cccn.2004.08.008.
Apolipoprotein B-48 (apoB-48) is produced by the small intestine, as a part of chylomicrons (CMs), and appears to be a suitable marker for clinical studies of postprandial lipoproteins and related cardiovascular risk factors. We have developed an assay for routine analysis to quantify apoB-48 in serum or plasma.
A microtiter plate was coated with monoclonal antibody (4C8) raised against apoB-48 C-terminal specific decapeptide. Serum samples were diluted 100-fold with 0.05 mol/l Tris-HCl buffer (with or without 0.1% Triton X-100). Appropriate calibration curves were obtained in the ELISA by using apoB-48 recombinant antigen.
No cross-reactivity (<0.001%) was found with apoB-100, as verified by ELISA and Western blot analyses. Intra- and inter-assay CVs were 4.8% and 5.4%, respectively. Recovery of added recombinant apoB-48 in serum was within 94-105%. The assay linearity was intact >5-fold dilution of serum by dilution buffer. ApoB-48 levels in healthy controls (n=18) at fasting were within the range of 2.69-6.56 microg/ml (mean+/-S.D.: 4.60+/-1.54 microg/ml). In healthy subjects, serum apoB-48 concentrations markedly increased in the postprandial state, in parallel with serum triglycerides.
This method for measuring apoB-48 using the monoclonal antibody 4C8 is simple, reliable and suitable for routine analyses.
载脂蛋白B-48（apoB-48）由小肠产生，是乳糜微粒（CMs）的一部分，似乎是餐后脂蛋白及相关心血管危险因素临床研究的合适标志物。我们已开发出一种用于常规分析的检测方法，以定量血清或血浆中的apoB-48。
用针对apoB-48 C末端特异性十肽的单克隆抗体（4C8）包被微量滴定板。血清样本用0.05 mol/l Tris-HCl缓冲液（含或不含0.1% Triton X-100）稀释100倍。通过使用apoB-48重组抗原在酶联免疫吸附测定（ELISA）中获得适当的校准曲线。
通过ELISA和蛋白质印迹分析验证，与apoB-100无交叉反应（<0.001%）。批内和批间变异系数分别为4.8%和5.4%。血清中添加的重组apoB-48回收率在94%至105%之间。该检测方法的线性在血清用稀释缓冲液稀释>5倍时仍保持完整。18名健康对照者空腹时的apoB-48水平在2.69 - 6.56 μg/ml范围内（平均值±标准差：4.60±1.54 μg/ml）。在健康受试者中，餐后状态下血清apoB-48浓度显著升高，与血清甘油三酯平行。
这种使用单克隆抗体4C8测量apoB-48的方法简单、可靠，适用于常规分析。
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