HOMOGENEOUS FERROCENE-MEDIATED AMPEROMETRIC IMMUNOASSAY
HOMOGENEOUS FERROCENE-MEDIATED AMPEROMETRIC IMMUNOASSAY
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DOI：
10.1021/ac00297a050
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发表时间：
1986-05-01
影响因子：
7.4
通讯作者：
GREEN, MJ
中科院分区：
文献类型：
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作者：
DIGLERIA, K;HILL, HAO;GREEN, MJ
本文报道了一种基于铁离子介导的葡萄糖氧化酶氧化葡萄糖的电流型免疫电极。本文研究了一种抗铁离子络合物对葡萄糖氧化酶的电子受体作用。当抗原-二茂铁复合物与抗体结合时，葡萄糖酶促氧化产生的催化电流被特异性抑制。加入游离抗原后，抑制作用可逆转。因此，设计了一种同质的竞争性免疫测定法。选择利多卡因作为抗原。测定时间为15 min，相对标准偏差为3- 6%。在过去的十年中，人们对开发用于测定临床和生理相关物质浓度的快速、简单的酶电极越来越感兴趣（7-3）。酶电极将生物催化剂对其底物的特异性和亲和力与电化学装置的分析能力相结合。当与化学放大机制（如酶催化）结合使用时，电化学方法的灵敏度和检测限得到改善（4）。电化学检测器与更广泛使用的分光光度技术相比有两个主要优点：（a）它们能够在混浊的样品中进行测量;（B）设备相对便宜且易于操作。对开发灵敏酶电极的兴趣已扩展到免疫化学领域，在该领域，开发非放射性同位素标记的免疫测定变得非常重要（5）。已经证明，在某些情况下，酶免疫测定可以具有与放射性同位素标记免疫测定相同的固有灵敏度（6）。应用免疫化学和电化学方法测量临床上重要的物种结合了酶电极的灵敏度和抗体-抗原相互作用的特异性。电化学免疫分析的一些应用-
An amperometric Immunoelectrode based on the ferrlcinlumlon-medlated oxidation of glucose by glucose oxidase Is described. An antlgen-ferridclnlum Ion complex was shown to act as an electron acceptor for glucose oxidase. The cata-lytic current produced In the enzymatic oxidation of glucose was specifically Inhibited upon binding the antigen-ferrocene complex with antibody. The Inhibition could be reversed upon addition of free antigen. Thus, a homogeneous, competitive Immunoassay was devised. Lidocaine was chosen as the antigen. The assay time was 15 min, with a relative standard deviation of 3-6%.Over the past decade there has been a growing interest in the development of rapid, simple enzyme electrodes for the determination of the concentration of clinically and physiologically relevant substances (7-3). An enzyme electrode combines the specificity and affinity of a biological catalyst for its substrate with the analytical power of electrochemical devices. The sensitivity and detection limits of electrochemical methods are improved when used in conjunction with chemical amplification mechanisms such as enzyme catalysis (4). Electrochemical detectors have two major advantages over the more widely used spectrophotometric techniques:(a) they are capable of making measurements in turbid samples and (b) the devices are relatively inexpensiveand easy to operate. The interest in developing sensitiveenzyme electrodes has expanded into the field of immunochemistry where it has become important to develop nonradioisotope-labeled im-munoassays (5). It has been demonstrated that enzyme im-munoassay can, in some cases, have the same intrinsic sen-sitivity as radioisotope-labeled immunoassay (6). The ap-plication of immunochemicaland electrochemical methods to the measurement of clinically important species combines the sensitivity of an enzyme electrode with the specificity of an antibody-antigen interaction. Some previousapplications of electrochemical immunoas-