Simple Determination of Trace Amounts of Antigen Using Autocatalytic Reaction with Na2SO3 and H2O2 Followed by Enzyme Immunoassay
Simple Determination of Trace Amounts of Antigen Using Autocatalytic Reaction with Na2SO3 and H2O2 Followed by Enzyme Immunoassay
复制标题
使用 Na2SO3 和 H2O2 自催化反应随后进行酶免疫分析简单测定痕量抗原
DOI：
10.1081/al-120017268
复制
发表时间：
2003
影响因子：
2
通讯作者：
S. Igarashi
中科院分区：
文献类型：
--
作者：
A. Manaka;S. Igarashi
过氧化氢氧化亚硫酸根离子为硫酸根离子是一个自催化反应，其中质子（H+）既作为反应产物又作为反应物。该反应提供了特征性的S形pH-时间曲线，该曲线包含拐点时间和终点处的动态pH变化。拐点时间随辣根过氧化物酶（HRP）浓度的变化而变化。此外，可以使用pH指示剂如溴百里酚蓝观察终点的pH变化。根据这些发现，使用秒表和肉眼，一个简单的和直观的测定mg/L水平的HRP已经开发。HRP的测定范围为0.2 ~ 50 mg/L，对空白样品8次测定的相对标准偏差为1.7%。检出限（3σ）为0.2mg/L。两种抗原（例如，以HRP为标记酶的酶免疫分析法与目视法相结合，可用于肿瘤坏死因子TNF-α和内分泌干扰物双酚A的检测。
Abstract The oxidation from sulfite ion to sulfate ion with hydrogen peroxide is an autocatalytic reaction, in which a proton (H+) acts not only as the reaction product but also as the reactant. This reaction provides the characteristic S-shaped pH vs. time curve, which contains the inflection time and the dynamic pH change at the end point. The inflection time shifts with the horseradish peroxidase (HRP) concentration. Furthermore, the pH change at the end point can be visualized using a pH indicator such as bromothymol blue. Based on these findings, using a stopwatch and the unaided eye, a simple and visual determination for mg/L levels of HRP has been developed. The determination range of HRP was from 0.2 to 50 mg/L, and the relative standard deviation was 1.7% (eight determinations) for the blank. The detection limits (3σ) were 0.2 mg/L. Two kinds of antigens (e.g., TNF-α as the tumor necrosis factor and bis-phenol A as the endocrine disrupting chemicals) were possible to determine by combination with the proposed visual technique and the enzyme immunoassay with HRP as a marker enzyme.