Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresisopen access
- Authors
- Lee, SangWook; Kim, Byung Woo; Shin, Hye-Su; Go, Anna; Lee, Min-Ho; Lee, Dong-Ki; Kim, Soyoun; Jeong, Ok Chan
- Issue Date
- Nov-2019
- Publisher
- Multidisciplinary Digital Publishing Institute (MDPI)
- Keywords
- aptamer; acoustophoresis; microfluidics; gram-negative bacteria
- Citation
- Micromachines, v.10, no.11
- Journal Title
- Micromachines
- Volume
- 10
- Number
- 11
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/37432
- DOI
- 10.3390/mi10110770
- ISSN
- 2072-666X
2072-666X
- Abstract
- 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 mu m) 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 mu L/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.
- Files in This Item
-
- Appears in
Collections - College of ICT Engineering > School of Integrative Engineering > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.