一种基于新型微芯片电泳的化学发光免疫分析方法，用于检测人血清中的甲胎蛋白。
A novel microchip electrophoresis-based chemiluminescence immunoassay for the detection of alpha-fetoprotein in human serum.
机构信息
State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection of Ministry Education, Guangxi Normal University, Guilin 541004, China.
State Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection of Ministry Education, Guangxi Normal University, Guilin 541004, China.
出版信息
Talanta. 2017 Apr 1;165:107-111. doi: 10.1016/j.talanta.2016.12.038. Epub 2016 Dec 19.
A sensitive immunoassay method based on microchip electrophoresis chemiluminescence (MCE-CL) detection technology was developed for the detection of tumor marker alpha-fetoprotein (AFP). This method adopts the non-competitive immunoassay mode, and was conducted after AFP reacted with excessive horseradish peroxidase (HRP) labeled monoclonal antibody. The extreme pH value was adopted in the electrophoresis buffer solution. The use of brij 35 as an additive of electrophoresis buffer increased dramatically the resolution (Rs) and the reproducibility of the analysis. Under the optimized experimental conditions, effective separation of the immune complex Ag-Ab* and free Ab* was achieved within 60s. The peak height of the immune complex Ag-Ab* was taken as quantification of AFP. Good linearity was observed within AFP concentrations ranging from 10ng/mL to 150ng/mL, and the detection limit was found to be 7.2ng/mL (1.0×10M). The present method was successfully applied for the detection of AFP in human serum from both healthy and cancer patients, and the AFP levels in the both were found be in the range of 16.5-23.4ng/mL and 416.2-825.4ng/mL, respectively.
基于微芯片电泳化学发光（MCE-CL）检测技术的灵敏免疫分析方法被开发用于检测肿瘤标志物甲胎蛋白（AFP）。该方法采用非竞争免疫分析模式，在 AFP 与过量辣根过氧化物酶（HRP）标记的单克隆抗体反应后进行。电泳缓冲溶液采用极端 pH 值。在电泳缓冲液中添加 Brij 35 可显著提高分析的分辨率（Rs）和重现性。在优化的实验条件下，在 60s 内实现了免疫复合物 Ag-Ab和游离 Ab的有效分离。免疫复合物 Ag-Ab*的峰高被用作 AFP 的定量。在 AFP 浓度范围为 10ng/mL 至 150ng/mL 内观察到良好的线性关系，检测限为 7.2ng/mL（1.0×10M）。本方法成功地应用于检测来自健康和癌症患者的人血清中的 AFP，并且在两者中发现 AFP 水平分别在 16.5-23.4ng/mL 和 416.2-825.4ng/mL 的范围内。
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