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Enzyme-linked immunoassay of alpha-1-fetoprotein in serum by differential pulse voltammetry

机译：差示脉冲伏安法测定血清中α-1-甲胎蛋白的酶联免疫法

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摘要

A method for the determination of alpha-1-fetoprotein (AFP) in human serum by using a horseradish peroxidase (HRP) label in an enzyme-linked immunosorbent assay (ELISA) is proposed. The method is based on the electrochemical determination of enzymatic reaction product with differential pulse voltammetry at a gold disk electrode. The assay consists of two successive steps. The first strip is a conventional HRP-mediated ELISA for the formation of electroactive 2,2'-diaminoazobenzene by means of the o-phenylenediamine-H2O2-HRP system. At the second step, 2.2'-diaminoazobenzene exhibits a sensitive voltammetric response at -0.19 V in pH 2.0 PBS. The peak current is proportional to the concentration of AFP in the range of 0.5-400 ng/mL (R = 0.9993) under optimum conditions. The sensitivity of this method is higher than that of the traditional spectrophotometric ELISA procedure. The proposed method has been applied to the clinical determination of AFP in human serum with satisfactory precision and accuracy. [References: 21]
机译：提出了一种通过辣根过氧化物酶（HRP）标记在酶联免疫吸附测定（ELISA）中测定人血清中α-1-甲胎蛋白（AFP）的方法。该方法基于在金盘电极上用差分脉冲伏安法电化学测定酶促反应产物。该测定法包括两个连续的步骤。第一条是常规的HRP介导的ELISA，用于通过邻苯二胺-H2O2-HRP系统形成电活性的2,2'-二氨基偶氮苯。在第二步中，在pH 2.0 PBS中，2.2'-二氨基偶氮苯在-0.19 V处显示出灵敏的伏安响应。在最佳条件下，峰值电流与AFP的浓度成正比，范围为0.5-400 ng / mL（R = 0.9993）。该方法的灵敏度高于传统的分光光度ELISA程序。该方法已用于临床测定人血清中AFP，具有良好的精密度和准确性。 [参考：21]

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