1. Targeted capture of pathogenic bacteria using a mammalian cell receptor coupled with dielectrophoresis on a biochip
- Author
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Koo, Ok Kyung, Liu, YiShao, Shuaib, Salamat, Bhattacharya, Shantanu, Ladisch, Michael R., Bashir, Rashid, and Bhunia, Arun K.
- Subjects
Bacteria, Pathogenic -- Physiological aspects ,Biosensors -- Research ,Cell receptors -- Properties ,Cell receptors -- Usage ,Mammals -- Physiological aspects ,Mammals -- Health aspects ,Microfluidics -- Research ,Fluidic devices -- Design and construction ,Fluidic devices -- Usage ,Electrophoresis -- Research ,Chemistry - Abstract
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.
- Published
- 2009