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在固定相芯片实验室中通过移动磁性颗粒进行麻痹性贝类毒素的免疫分析。

Immunoassay of paralytic shellfish toxins by moving magnetic particles in a stationary liquid-phase lab-on-a-chip.

机构信息

Department of Chemistry, Gangneung-Wonju National University, Gangneung 210-702, Republic of Korea.

Department of Chemistry, Gangneung-Wonju National University, Gangneung 210-702, Republic of Korea.

出版信息

Biosens Bioelectron. 2015 Apr 15;66:136-40. doi: 10.1016/j.bios.2014.11.012. Epub 2014 Nov 11.

Abstract

In this study, we devised a stationary liquid-phase lab-on-a-chip (SLP LOC), which was operated by moving solid-phase magnetic particles in the stationary liquid phase. The SLP LOC consisted of a sample chamber to which a sample and reactants were added, a detection chamber containing enzyme substrate solution, and a narrow channel connecting the two chambers and filled with buffer. As a model system, competitive immunoassays of saxitoxin (STX), a paralytic shellfish toxin, were conducted in the SLP LOC using protein G-coupled magnetic particles (G-MPs) as the solid phase. Anti-STX antibodies, STX-horseradish peroxidase conjugate, G-MPs, and a STX sample were added to the sample chamber and reacted by shaking. While liquids were in the stationary state, G-MPs were transported from the sample chamber to the detection chamber by moving a magnet below the LOC. After incubation to allow the enzymatic reaction to occur, the absorbance of the detection chamber solution was found to be reciprocally related to the STX concentration of the sample. Thus, the SLP LOC may represent a novel, simple format for point-of-care testing applications of enzyme-linked immunosorbent assays by eliminating complicated liquid handling steps.

摘要

在这项研究中,我们设计了一种固定相液相微流控芯片(SLP LOC),它通过在固定相液相中移动固相磁性颗粒来操作。SLP LOC 由一个样品腔组成,向其中添加样品和反应物,一个检测腔含有酶底物溶液,以及一个连接两个腔并充满缓冲液的狭窄通道。作为模型系统,使用蛋白 G 偶联磁性颗粒(G-MP)作为固相,在 SLP LOC 中进行了石房蛤毒素(STX)的竞争性免疫测定。将抗 STX 抗体、STX-辣根过氧化物酶缀合物、G-MP 和 STX 样品添加到样品腔中,并通过摇动进行反应。当液体处于静止状态时,通过在 LOC 下方移动磁铁将 G-MP 从样品腔输送到检测腔。孵育以允许酶反应发生后,发现检测腔溶液的吸光度与样品中 STX 的浓度呈反比关系。因此,通过消除复杂的液体处理步骤,SLP LOC 可能代表酶联免疫吸附测定用于即时检测应用的一种新颖、简单的格式。

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