Targeted Capture of Pathogenic Bacteria Using a Mammalian Cell Receptor Coupled with Dielectrophoresis on a Biochip
Targeted Capture of Pathogenic Bacteria Using a Mammalian Cell Receptor Coupled with Dielectrophoresis on a Biochip
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DOI：
10.1021/ac9000833
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发表时间：
2009-04-15
影响因子：
7.4
通讯作者：
Bhunia, Arun K.
中科院分区：
文献类型：
--
作者：
Koo, Ok Kyung;Liu, YiShao;Bhunia, Arun K.
在生物传感器平台上有效捕获目标分析物是在微流控芯片中可靠和特异性检测病原微生物的先决条件。抗体已被广泛用作配体;然而，由于其偶尔的不令人满意的性能，正在寻找替代受体。热休克蛋白60（Heat shock protein 60，Hsp 60）是单核细胞增生李斯特菌（Listeria monocytogenes）感染过程中粘附蛋白（Listeria adhesion protein，HSP 60）的受体。本文报道了生物素化的Hsp 60作为活的（活的）单核细胞增多性李斯特菌在微流体环境中的捕获分子的应用。Hsp 60固定在链霉亲和素包被的二氧化硅的表面上，表现出针对其他李斯特菌属物种、沙门氏菌、埃希氏菌、芽孢杆菌、假单胞菌、沙雷氏菌、哈夫尼氏菌、肠杆菌、柠檬酸杆菌和乳杆菌的背景的致病性李斯特菌的特异性捕获。单核细胞增生李斯特菌的捕获效率是另一种李斯特菌受体单克隆抗体mAb-C11 E9的83倍。此外，捕获率进一步增加了60%的热休克蛋白60包被的生物芯片时，在最后的1小时孵育步骤开始时，施加5分钟的介电泳力。表明Hsp 60可用于L.在生物芯片传感器平台上检测单核细胞增多症。
Efficient capture of target analyte on biosensor platforms is a prerequisite for reliable and specific detection of pathogenic microorganisms in a microfluidic chip. Antibodies have been widely used as ligands; however, because of their occasional unsatisfactory performance, a search for alternative receptors is underway. Heat shock protein 60 (Hsp60), a eukaryotic mitochondrial chaperon protein is a receptor for Listeria adhesion protein (LAP) during Listeria monocytogenes infection. This paper reports application of biotinylated Hsp60 as a capture molecule for living (viable) L monocytogenes in a microfluidic environment Hsp60, immobilized on the surface of streptavidin-coated silicon dioxide exhibited specific capture of pathogenic Listeria against a background of other Listeria species, Salmonella, Escherichia, Bacillus, Pseudomonas, Serratia, Hafnia, Enterobacter, Citrobacter, and Lactobacillus. The capture efficiency of L monocytogenes was 83 times greater than another Listeria receptor, the monoclonal antibody, mAb-C11E9. Additionally, the capture rate was further increased on a Hsp60-coated biochip by 60% when a dielectrophoresis force was applied for 5 min at the beginning of the final 1 h incubation step. Our data show that Hsp60 could be used for specific detection of L. monocytogenes on a biochip sensor platform.