基于毛细管电泳的牛初乳产品中免疫球蛋白G的非竞争性免疫分析

CE-based noncompetitive immunoassay for immunoglobulin G in bovine colostrum products.

作者信息

Zhao Jin, Ding Xiaojing, Wang Xinyu, Wang Qian, Wang Zhi

机构信息

Beijing Center for Disease Prevention and Control, Beijing, P. R. China.

出版信息

Electrophoresis. 2007 Nov;28(21):3934-9. doi: 10.1002/elps.200700122.

DOI:10.1002/elps.200700122

PMID:17922500

Abstract

A CE-based noncompetitive immunoassay for IgG in bovine colostrum products was established. FITC-labeled protein G (FITC-PrG) was tagged through noncovalent bindings to the Fc region of the mouse monoclonal antibovine IgG (Ab). The FITC-PrG, Ab, and IgG formed a sandwiched immunocomplex FITC-PrG-Ab-IgG under optimal incubation conditions. The immunocomplex was separated and analyzed by CZE with LIF detection in less than 2 min in an uncoated fused-silica capillary. Addition of PEG 20,000 (PEG 20M) in the running buffer significantly suppressed analyte adsorption and thus improved the reproducibility and the resolution. The precision of the method was 5.1% (n = 7). A linear relationship was established for the IgG concentration in the range of 1-5 mg/L with a linear correlation coefficient (r = 0.9917). The LOD was 0.1 mg/L (S/N = 3). The method was successfully applied for the determination of IgG in bovine colostrum products and satisfactory results were achieved.

摘要

建立了一种基于毛细管电泳的牛初乳产品中IgG的非竞争性免疫分析方法。异硫氰酸荧光素标记的蛋白G（FITC-PrG）通过非共价结合标记到小鼠抗牛IgG单克隆抗体（Ab）的Fc区域。在最佳孵育条件下，FITC-PrG、Ab和IgG形成夹心免疫复合物FITC-PrG-Ab-IgG。在未涂层的熔融石英毛细管中，不到2分钟即可通过毛细管区带电泳结合激光诱导荧光检测对免疫复合物进行分离和分析。在运行缓冲液中添加聚乙二醇20000（PEG 20M）可显著抑制分析物吸附，从而提高重现性和分辨率。该方法的精密度为5.1%（n = 7）。在1-5 mg/L范围内建立了IgG浓度的线性关系，线性相关系数（r = 0.9917）。检测限为0.1 mg/L（信噪比 = 3）。该方法成功应用于牛初乳产品中IgG的测定，获得了满意的结果。