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用于同时检测胃蛋白酶原1和2的具有可分离电极和抗体芯片的电化学微器件。

Electrochemical microdevice with separable electrode and antibody chips for simultaneous detection of pepsinogens 1 and 2.

作者信息

Ogasawara Daichi, Hirano Yu, Yasukawa Tomoyuki, Shiku Hitoshi, Kobori Kiichirou, Ushizawa Koji, Kawabata Shohei, Matsue Tomokazu

机构信息

Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aoba, Aramaki, Sendai 980-8579, Japan.

出版信息

Biosens Bioelectron. 2006 Mar 15;21(9):1784-90. doi: 10.1016/j.bios.2005.09.009. Epub 2005 Oct 21.

Abstract

An electrochemical microdevice with separable electrode and antibody chips has been developed and applied to detect atrophic gastritis-related proteins, pepsinogen 1 (PG1) and pepsinogen 2 (PG2), based on sandwich-type enzyme-linked immunosorbent assays (ELISAs) with horseradish peroxidase (HRP)-labeled antibody. To fabricate the electrochemical device for simultaneous analysis of several proteins, the electrode chip with eight electrode elements was assembled along with an antibody chip with eight cavities containing immobilized anti-PG1 or anti-PG2. The immunoreactions occurring in the cavities of the device were detected simultaneously by amperometry. The labeled HRP in the cavity in the presence of hydrogen peroxide catalyzed the oxidation of ferrocenemethanol (FMA) to FMA+, which was detected electrochemically by the electrode chip. The amperometric responses of respective cavities in the device increased with increasing concentration of PG1 or PG2 of 0-50 ng/ml, ensuring the simultaneous detection of PG1 and PG2. The detection limits for both PG1 and PG2 were 0.6 ng/ml (S/N=2). The electrode chip was recovered easily by disassembling the electrochemical device; thereby, it was used repeatedly, whereas the antibody chip was discarded. No marked decrease in electrochemical responses was detected after repeated use. Reuse of the electrode chip is beneficial to reduce costs of protein analysis.

摘要

一种具有可分离电极和抗体芯片的电化学微器件已被开发出来,并基于用辣根过氧化物酶(HRP)标记抗体的夹心型酶联免疫吸附测定(ELISA)用于检测萎缩性胃炎相关蛋白胃蛋白酶原1(PG1)和胃蛋白酶原2(PG2)。为了制造用于同时分析多种蛋白质的电化学器件,将具有八个电极元件的电极芯片与一个具有八个腔室的抗体芯片组装在一起,这些腔室中固定有抗PG1或抗PG2。通过安培法同时检测器件腔室中发生的免疫反应。在过氧化氢存在下,腔室中的标记HRP催化二茂铁甲醇(FMA)氧化为FMA +,电极芯片通过电化学方法检测到FMA +。随着PG1或PG2浓度在0 - 50 ng/ml范围内增加,器件中各个腔室的安培响应增加,从而确保了PG1和PG2的同时检测。PG1和PG2的检测限均为0.6 ng/ml(S/N = 2)。通过拆卸电化学器件可轻松回收电极芯片;因此,电极芯片可重复使用,而抗体芯片则被丢弃。重复使用后未检测到电化学响应有明显下降。电极芯片的重复使用有利于降低蛋白质分析成本。

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