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使用单克隆抗体对IgG进行比浊测定。

Turbidimetric assay of IgG with use of single monoclonal antibodies.

作者信息

Maynard Y, Scott M G, Nahm M H, Ladenson J H

出版信息

Clin Chem. 1986 May;32(5):752-7.

PMID:3698266
Abstract

We describe a turbidimetric assay for quantifying total immunoglobulin G (IgG) in serum with use of a single monoclonal antibody. The reaction, monitored by a centrifugal analyzer, is technically simple, rapid, and precise. Buffer of low ionic strength and polyethylene glycol are required for formation of detectable antibody-antigen complexes. We measured IgG concentrations in 49 polyclonal sera (Group 1) and 84 sera containing monoclonal IgG (Group 2) in assays in which we used either of two anti-IgG monoclonal antibodies (HG6 or HG8). Results compared well with those obtained with a nephelometric assay involving polyclonal antiserum, except for sera from four persons of Group 2 whose immunoglobulins were not detected by antibody HG6. HG6 bound IgG from these four sera in a solid-phase binding assay. HG6 and HG8 recognize epitopes on the Fab and Fc regions of IgG, respectively, and they do not compete for binding to the whole molecule. However, use of the two monoclonal antibodies combined failed to improve the sensitivity or range of the assay. We conclude that light-scattering assays of IgG can be validly performed with a single monoclonal antibody.

摘要

我们描述了一种使用单克隆抗体定量血清中总免疫球蛋白G(IgG)的比浊法。该反应由离心分析仪监测,技术上简单、快速且精确。形成可检测的抗体-抗原复合物需要低离子强度的缓冲液和聚乙二醇。在使用两种抗IgG单克隆抗体(HG6或HG8)之一的测定中,我们测量了49份多克隆血清(第1组)和84份含单克隆IgG的血清(第2组)中的IgG浓度。除了第2组中4人的血清未被抗体HG6检测到免疫球蛋白外,结果与使用多克隆抗血清的散射比浊法所获得的结果相比良好。HG6在固相结合试验中能结合这4份血清中的IgG。HG6和HG8分别识别IgG Fab和Fc区域上的表位,它们不会竞争与整个分子的结合。然而,联合使用这两种单克隆抗体未能提高该测定的灵敏度或范围。我们得出结论,使用单克隆抗体可以有效地进行IgG的光散射测定。

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