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Open AccessArticle

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

1
PCL Incorporated, Seoul 08510, Korea
2
Institute of Digital Anti-Aging Health Care, Inje University, Gimhea 50834, Korea
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Department of Chemistry, Sungyunkwan University, Suwon 16419, Korea
4
School of Integrative Engineering, Chung-Ang University, Seoul 06974, Korea
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Department of Biomedical Engineering, Dongguk University, Seoul 10326, Korea
6
Department of Biomedical Engineering, Inje University, Gimhea 50834, Korea
*
Authors to whom correspondence should be addressed.
Micromachines 2019, 10(11), 770; https://doi.org/10.3390/mi10110770
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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