利用单克隆抗体对血浆中载脂蛋白AII进行酶联免疫吸附测定。

Enzyme-linked immunoabsorbant assay of apolipoprotein AII in plasma, with use of a monoclonal antibody.

作者信息

Stein E A, DiPersio L, Pesce A J, Kashyap M, Kao J T, Srivastava L, McNerney C

出版信息

Clin Chem. 1986 Jun;32(6):967-71.

Abstract

We produced a monoclonal antibody (C2-22) to human apolipoprotein (Apo) AII and describe its use in an enzyme-linked immunoabsorbant assay (ELISA) for Apo AII in human plasma and lipoprotein subfractions. No cross reactivity of the antibody with Apo CI, CII, CIII, E, or ablumin was detected. Apo AI and low- and very-low-density lipoprotein cross reacted by 0.25%, less than 0.2%, and less than 0.3%, respectively. Whole plasma high-density lipoprotein (HDL) and HDL subfractions (HDL2 and HDL3) produced parallel displacement curves. This quantitative ELISA is based on competition between solid-phase-bound Apo AII and free Apo AII. Bound C2-22 is detected by alkaline-phosphatase-labeled second antibody. The standard curve for the assay is linear for plasma diluted 500-fold originally containing 140 to 1140 mg of Apo AII per liter. Delipidation of plasma samples exposed no additional antigenic sites. Within- and between-run CVs were respectively 8.4% and 8.7% at 327 mg/L of Apo AII, and 6.8% and 7.4% at 587 mg/L. Results correlated well with those by a polyvalent-antisera-based RIA procedure: r = 0.916, p less than 0.01, RIA = 0.896 ELISA -19.1 mg/L.

摘要

我们制备了一种针对人载脂蛋白（Apo）AII的单克隆抗体（C2 - 22），并描述了其在酶联免疫吸附测定（ELISA）中用于检测人血浆和脂蛋白亚组分中Apo AII的用途。未检测到该抗体与Apo CI、CII、CIII、E或白蛋白有交叉反应。Apo AI与低密度脂蛋白和极低密度脂蛋白的交叉反应率分别为0.25%、小于0.2%和小于0.3%。全血浆高密度脂蛋白（HDL）及其亚组分（HDL2和HDL3）产生平行的置换曲线。这种定量ELISA基于固相结合的Apo AII与游离Apo AII之间的竞争。通过碱性磷酸酶标记的二抗检测结合的C2 - 22。该测定的标准曲线对于最初稀释500倍、每升含140至1140毫克Apo AII的血浆呈线性。血浆样品的脱脂未暴露额外的抗原位点。在Apo AII浓度为327毫克/升时，批内和批间变异系数分别为8.4%和8.7%；在587毫克/升时，分别为6.8%和7.4%。结果与基于多价抗血清的放射免疫分析（RIA）程序的结果相关性良好：r = 0.916，p小于0.01，RIA = 0.896 ELISA - 19.1毫克/升。