Schmalzing D, Nashabeh W, Yao X W, Mhatre R, Regnier F E, Afeyan N B, Fuchs M
PerSeptive Biosystems, Cambridge, Massachusetts 02139.
Anal Chem. 1995 Feb 1;67(3):606-12. doi: 10.1021/ac00099a019.
A competitive immunoassay for cortisol based on capillary electrophoresis (CE) and laser-induced fluorescence is described. The work involved the production of assay reagents and the development of separation conditions allowing for routine analysis of serum samples. Fluorescein-labeled cortisol was synthesized and purified. Fab fragments were produced from mouse monoclonal anticortisol antibody and purified using a POROS cation exchange chromatography column. After incubation of these reagents with serum, free and bound labeled antigen were separated by CE with high reproducibility. No prior sample cleanup of the serum samples was necessary. Serum calibration curves were established and used for the quantitation of cortisol in serum. The results demonstrate feasibility for a cortisol assay based on CE operating directly on serum samples.
描述了一种基于毛细管电泳（CE）和激光诱导荧光的皮质醇竞争性免疫测定法。这项工作包括测定试剂的制备以及分离条件的优化，以实现血清样本的常规分析。合成并纯化了荧光素标记的皮质醇。从小鼠单克隆抗皮质醇抗体中制备Fab片段，并使用POROS阳离子交换色谱柱进行纯化。将这些试剂与血清孵育后，通过CE可高度重现地分离游离和结合的标记抗原。血清样本无需预先进行净化处理。建立了血清校准曲线并用于血清中皮质醇的定量分析。结果表明基于CE直接对血清样本进行操作的皮质醇测定法具有可行性。
