Preprint Article Version 1 This version is not peer-reviewed

Aptamer Bead-Mediated Separation of Gram-Negative Bacteria Using Acoustophoresis

Version 1 : Received: 17 October 2019 / Approved: 18 October 2019 / Online: 18 October 2019 (11:41:41 CEST)

A peer-reviewed article of this Preprint also exists.

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. 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.

Journal reference: Micromachines 2019, 10, 770
DOI: 10.3390/mi10110770

Abstract

Here, we report a simple and effective method for separation 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. Using acoustophoresis, 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), and a concentration factor of up to 20×. The acoustophoresis microfluidic device also showed binding affinity to multiple strains of gram-negative bacteria, but not to gram-positive bacteria. This study presents a new paradigm for early diagnosis of 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.

Subject Areas

aptamer: acoustophoresis; microfluidics; gram-negative bacteria

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