分析化学2009，Vol.37Issue(7)：994-998,5.
2-氨基-3-羟基吡啶-过氧化氢-辣根过氧化物酶伏安酶联免疫体系分析人血清癌胚抗原
2-Amino-3-hydroxylpyridinE-H2O2-Horseradish Peroxidase Voltammetric EnzymE-Linked Immunoassay for Detection of Carcionembryonic Antigen in Human Serum
摘要
Abstract
2-Amino-3-hydroxylpyridinE-H2O2-horseradish peroxidase (HRP) voltammetric enzymE-linked immunoassay based on N-heterocyclic substrate has been successfully applied for the detection of carcionembryonic antigen(CEA) in human serum. 2-Amino-3-hydroxylpyridine is oxidized with H2O2 catalyzed by HRP, and the resulting electroactive product produces a sensitive voltammetric peak at the potential of 0.36 V(vs. SCE) in Britton-Robinson (B-R) buffer solution. By this voltammetric peak, free HRP can be measured and immunoassay of HRP label can be developed. The enzymE-catalyzed reaction conditions and voltammetric detection conditions have been optimized, and the electrode procedure of the enzymatic product was investigated. The selected optimum reaction conditions were that the reaction medium was pH 6.0 B-R buffer solution for 10 mL reaction solution containing 1.0 mL of 0.2 mol/L B-R buffer solution, 3.0 mL of 8.0 mmol/L 2-amino-3-hydroxylpyridine solution and 1.5 mL of 0.5 mmol/L H2O2 solution, and the reaction time was 30 min at 37 ℃. The optimum detection conditions were that the supporting electrolyte was pH 7.0 B-R buffer solution for 10 mL of the overall detection solution containing 5 mL of reaction solution and 1.0 mL of 0.2 mol/L B-R buffer solution. The optimum instrumental conditions for the detection were chosen as follows: the initial potential, 0.00 V; the final potential, -0.80 V; the potential scanning rate, 400 mV/s; the mercury drop standing time, 7 s. Under the optimum conditions, the linear range for detection of free HRP was 4.0×10-4-1.0 μg/L with a detection limit of 0.12 μg/L. Based on the new immunoassay system, the linear range of the detection to CEA was 0.50-80 μg/L and the detection limit was 0.50 μg/L, which is 10 times lower than that of traditional spectrophotometric enzymE-linked immunosorbent assay method. The 2-Amino-3-hydroxylpyridinE-H2O2-HRP voltammetric enzymE-linked immunoassay new system exhibits excellent performance having wider linear range and lower detection limit. The new method is inexpensive and simple. It has a great potential in clinical diagnosis.关键词
2-氨基-3-羟基吡啶/辣根过氧化物酶/伏安酶联免疫分析/癌胚抗原Key words
2-Amino-3-hydroxylpyridine, horseradish peroxidase, voltammetric enzymE-linked immunoassay, carcionembryonic antigen分类
化学化工引用本文复制引用
于凤丽,邹晋,梅振华..2-氨基-3-羟基吡啶-过氧化氢-辣根过氧化物酶伏安酶联免疫体系分析人血清癌胚抗原[J].分析化学,2009,37(7):994-998,5.基金项目
本文系国家自然科学基金(No.20775038)和青岛科技大学博士启动基金(No.0022297)资助项目 （No.20775038）
