Application of an automated fluidic system using electrochemical bead-based immunoassay to detect the bacteriophage MS2 and ovalbumin
Application of an automated fluidic system using electrochemical bead-based immunoassay to detect the bacteriophage MS2 and ovalbumin
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DOI：
10.1016/j.aca.2006.01.017
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发表时间：
2006-03-02
影响因子：
6.2
通讯作者：
Heineman, WR
中科院分区：
文献类型：
--
作者：
Kuramitz, H;Dziewatkoski, M;Heineman, WR
研制了一种用于电化学检测的全自动微珠免疫分析系统。使用该系统证明了对噬菌体MS 2和卵清蛋白（OVA）的测定。链霉亲和素包被的顺磁性微珠用作移动的固相。通过将生物素化的抗体连接到链霉亲和素包被的珠上，捕获抗原，然后将抗原暴露于与β-半乳糖苷酶缀合的抗体，制备免疫测定夹心。β-半乳糖苷酶将对氨基苯基吡喃半乳糖苷（PAPG）转化为对氨基苯酚（PAP），PAP被电化学氧化为顶级醌亚胺（PQI）。使用具有固定化β-半乳糖苷酶的珠粒，在不同的珠粒浓度和流速下，针对测定中的每个程序，研究了流体系统中顺磁性微珠的行为。此外，使用流体系统证明了全自动MS 2和OVA测定。MS 2和OVA的检测限分别为990（1.6 x 10（11）个颗粒/mL（-1））和470 ng/mL（-1）。(c)2006 Elsevier B. V.保留所有权利。
A fully-automated fluidic system for a bead-based immunoassay with electrochemical detection was developed. Assays for the bacteriophage MS2 and ovalbumin (OVA) were demonstrated using this system. Streptavidin-coated paramagnetic microbeads were used as a mobile solid phase. The immunoassay sandwich was made by attaching a biotinylated antibody to the streptavidin-coated beads, capturing antigen, and then exposing the antigen to an antibody conjugated with P-galactosidase. P-Galactosidase converts p-aminophenyl galactopyranoside (PAPG) to p-aminophenol (PAP), which is electrochemically oxidized top-quinone imine (PQI). The behavior of paramagnetic microbeads in the fluidic system was investigated using beads with immobilized P-galactosidase at different concentrations of beads and flow rates for each procedure in the assay. Furthermore, the fully-automated MS2 and OVA assays were demonstrated using the fluidic system. The limits of detection for MS2 and OVA were 990 (1.6 x 10(11) particles mL(-1)) and 470 ng mL(-1), respectively. (c) 2006 Elsevier B.V. All rights reserved.