Flow injection renewable surface immunoassay: a new approach to immunoanalysis with fluorescence detection.
Flow injection renewable surface immunoassay: a new approach to immunoanalysis with fluorescence detection.
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流动注射可再生表面免疫测定：一种带有荧光检测的免疫分析新方法。
DOI：
10.1021/ac00083a008
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发表时间：
1994
影响因子：
7.4
通讯作者：
Ruzicka,J
中科院分区：
文献类型：
--
作者：
Pollema,CH;Ruzicka,J
本文介绍了一种在可再生表面上使用流动注射技术自动进行非均相免疫分析的新方法。流动注射可再生表面免疫测定（FIRSI）依赖于使用微量珠粒来形成反应性表面，其通过荧光光谱法进行询问。在测定之后，通过流体去除用过的反应性表面并用新的珠层替换它，避免了通常用于基于流动的免疫测定的在线再生。这允许在早期阶段监测抗体-抗原结合，显著增加连续测定的采样频率，并消除与重复使用引起的表面反应性降低相关的问题。一个模型系统，利用抗小鼠IgG 1包被的珠和小鼠IgG 1蛋白被用来表征方法的再现性，流速，接触时间，和珠的量。异质性测定包括目前进行的免疫测定的大部分，1因为同质性测定的选择是有限的限制性要求，在结合时必须发生一些信号的变化。固相的使用通过物理分离分析物的结合和未结合部分来消除该要求，通常借助于固定在固相表面上的适当抗体。以这种方式，分析物选择性地结合到固定的抗体，而样品的剩余部分可以通过洗涤步骤去除。
This paper introduces a new methodology of carryingout heterogeneous immunoassays automatically, using a flow injection technique on a renewable surface. Flowinjection renewable surface immunoassay (FIRSI) relies on the use of a minute amount of beads to form a reactive surface, which is interrogated by fluorescence spectrometry. Following the assay, on-line regenerationnormally used in flow based immunoassays is avoided by fluidically removing the spent reactive surface and replacing it with a new layer of beads. This allows the monitoring of antibody-antigen binding at its early stages, dramatically increases the sampling frequency of a serial assay, and eliminates the problems associated with a decrease in surface reactivity caused by repetitive use. A model system utilizing anti-mouse IgGl-coated beads and mouse IgGl protein is used to characterize the method with respect to reproducibility, flow rate, contact time, and amount of beads.Heterogeneous assays comprise a majority of currently performed immunoassays, 1since the choice of homogeneous assays is limited by the restrictive requirement that some change in signal must occur upon binding. Use of a solid phase removes this requirement by physically separating the bound and the unbound portion of the analyte, typically by means of an appropriate antibody immobilized on the surface of a solid phase. In this way, the analyte is selectively bound to the immobilized antibody while the remaining portion of the sample can be removed by the washing steps.