Wang Q, Luo G, Ou J, Yeung W S
Department of Chemistry, Tsinghua University, Beijing, China.
J Chromatogr A. 1999 Jul 2;848(1-2):139-48. doi: 10.1016/s0021-9673(99)00413-6.
A new and simple approach to perform immunoassay using protein G affinity capillary chromatography and laser-induced fluorescence detection was described. A noncompetitive assay for monoclonal anti-bovine serum albumin (BSA) was used to test the performance of the system. Fluorescein isothiocyanate labeled BSA was used as a tracer to determine anti-BSA in pM level. Capillaries with inner diameter of 150 microns were packed with recombinant protein G-bound perfusive support. The packed capillary was used to capture the immunocomplexes, which were subsequently desorbed by 100 mM glycine (pH 9.0). Open tube capillary electrophoresis-based immunoassay (CEIA) for anti-BSA was also performed. Using standard samples, calibration curves for anti-BSA was established in both assays. Compared with CEIA, this system improved the concentration sensitivity for about 100-fold due to the pre-concentration of immunocomplex in the protein G column, while the mass sensitivity was similar in the two methods.
描述了一种使用蛋白G亲和毛细管色谱法和激光诱导荧光检测进行免疫分析的新的简单方法。采用非竞争性单克隆抗牛血清白蛋白（BSA）检测法来测试该系统的性能。用异硫氰酸荧光素标记的BSA作为示踪剂，以测定皮摩尔水平的抗BSA。内径为150微米的毛细管填充有结合重组蛋白G的灌注载体。填充毛细管用于捕获免疫复合物，随后用100 mM甘氨酸（pH 9.0）将其解吸。还进行了基于开管毛细管电泳的抗BSA免疫分析（CEIA）。使用标准样品，在两种检测方法中均建立了抗BSA的校准曲线。与CEIA相比，由于免疫复合物在蛋白G柱中的预浓缩，该系统将浓度灵敏度提高了约100倍，而两种方法的质量灵敏度相似。
