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Open AccessArticle

Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis

PCL Incorporated, Seoul 08510, Korea
Institute of Digital Anti-Aging Health Care, Inje University, Gimhea 50834, Korea
Department of Chemistry, Sungyunkwan University, Suwon 16419, Korea
School of Integrative Engineering, Chung-Ang University, Seoul 06974, Korea
Department of Biomedical Engineering, Dongguk University, Seoul 10326, Korea
Department of Biomedical Engineering, Inje University, Gimhea 50834, Korea
Authors to whom correspondence should be addressed.
Micromachines 2019, 10(11), 770;
Received: 17 October 2019 / Revised: 5 November 2019 / Accepted: 9 November 2019 / Published: 11 November 2019
Here, we report a simple and effective method for capturing and displacement of gram-negative bacteria using aptamer-modified microbeads and acoustophoresis. As acoustophoresis allows for simultaneous washing and size-dependent separation in continuous flow mode, we efficiently obtained gram-negative bacteria that showed high affinity without any additional washing steps. The proposed device has a simple and efficient channel design, utilizing a long, square-shaped microchannel that shows excellent separation performance in terms of the purity, recovery, and concentration factor. Microbeads (10 µm) coated with the GN6 aptamer can specifically bind gram-negative bacteria. After incubation of bacteria culture sample with aptamer affinity bead, gram-negative bacteria-bound microbeads, and other unbound/contaminants can be separated by size with high purity and recovery. The device demonstrated excellent separation performance, with high recovery (up to 98%), high purity (up to 99%), and a high-volume rate (500 µL/min). The acoustophoretic separation performances were conducted using 5 Gram-negative bacteria and 5 Gram-positive bacteria. Thanks to GN6 aptamer’s binding affinity, aptamer affinity bead also showed binding affinity to multiple strains of gram-negative bacteria, but not to gram-positive bacteria. GN6 coated bead can capture Gram-negative bacteria but not Gram-positive bacteria. This study may present a different perspective in the field of early diagnosis in bacterial infectious diseases. In addition to detecting living bacteria or bacteria-derived biomarkers, this protocol can be extended to monitoring the contamination of water resources and may aid quick responses to bioterrorism and pathogenic bacterial infections. View Full-Text
Keywords: aptamer; acoustophoresis; microfluidics; gram-negative bacteria aptamer; acoustophoresis; microfluidics; gram-negative bacteria
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Lee, S.; Kim, B.W.; Shin, H.-S.; Go, A.; Lee, M.-H.; Lee, D.-K.; Kim, S.; Jeong, O.C. Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis. Micromachines 2019, 10, 770.

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