Ou J P, Wang Q G, Cheung T M, Chan S T, Yeung W S
Department of Obstetrics and Gynaecology, The University of Hong Kong, Queen Mary Hospital, Hong Kong.
J Chromatogr B Biomed Sci Appl. 1999 Apr 30;727(1-2):63-71. doi: 10.1016/s0378-4347(99)00082-1.
A systematic study on the optimization of capillary electrophoresis-based immunoassay (CEIA) was performed using bovine serum albumin (BSA) and monoclonal anti-BSA. The immunocomplex could not be resolved from free BSA or anti-BSA with UV detection. When fluorescein isothiocyanate-labeled BSA (FITC-BSA) was used as tracer, the free and bound FITC-BSA were well separated giving definite peaks with laser induced fluorescence detection. The factors affecting the separation of the free and bound FITC-BSA, including voltage, pH and ionic strength of the running buffer, were systematically analyzed. Competitive CEIAs were demonstrated in uncoated and coated capillaries with whole or Fab fragment of the antibody. The coefficient of variation for the quantification of BSA in coated capillary was less than that in uncoated capillary. This study demonstrated that competitive CEIA could be applied to quantify high-molecular-mass protein in biological fluids.
使用牛血清白蛋白（BSA）和抗牛血清白蛋白单克隆抗体，对基于毛细管电泳的免疫分析（CEIA）优化进行了系统研究。通过紫外检测，免疫复合物无法与游离的BSA或抗BSA分离。当使用异硫氰酸荧光素标记的BSA（FITC-BSA）作为示踪剂时，游离和结合的FITC-BSA通过激光诱导荧光检测得到了很好的分离，给出了明确的峰。系统分析了影响游离和结合FITC-BSA分离的因素，包括运行缓冲液的电压、pH值和离子强度。在未涂层和涂层毛细管中，使用抗体的完整或Fab片段进行了竞争性CEIA。涂层毛细管中BSA定量的变异系数低于未涂层毛细管。本研究表明，竞争性CEIA可用于定量生物流体中的高分子质量蛋白质。
