Cargando…
Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis
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-negativ...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6915462/ https://www.ncbi.nlm.nih.gov/pubmed/31718045 http://dx.doi.org/10.3390/mi10110770 |
_version_ | 1783480021734653952 |
---|---|
author | Lee, SangWook Kim, Byung Woo Shin, Hye-Su Go, Anna Lee, Min-Ho Lee, Dong-Ki Kim, Soyoun Jeong, Ok Chan |
author_facet | Lee, SangWook Kim, Byung Woo Shin, Hye-Su Go, Anna Lee, Min-Ho Lee, Dong-Ki Kim, Soyoun Jeong, Ok Chan |
author_sort | Lee, SangWook |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6915462 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-69154622019-12-24 Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis Lee, SangWook Kim, Byung Woo Shin, Hye-Su Go, Anna Lee, Min-Ho Lee, Dong-Ki Kim, Soyoun Jeong, Ok Chan Micromachines (Basel) Article 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. MDPI 2019-11-11 /pmc/articles/PMC6915462/ /pubmed/31718045 http://dx.doi.org/10.3390/mi10110770 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Lee, SangWook Kim, Byung Woo Shin, Hye-Su Go, Anna Lee, Min-Ho Lee, Dong-Ki Kim, Soyoun Jeong, Ok Chan Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title | Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title_full | Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title_fullStr | Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title_full_unstemmed | Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title_short | Aptamer Affinity-Bead Mediated Capture and Displacement of Gram-Negative Bacteria Using Acoustophoresis |
title_sort | aptamer affinity-bead mediated capture and displacement of gram-negative bacteria using acoustophoresis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6915462/ https://www.ncbi.nlm.nih.gov/pubmed/31718045 http://dx.doi.org/10.3390/mi10110770 |
work_keys_str_mv | AT leesangwook aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT kimbyungwoo aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT shinhyesu aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT goanna aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT leeminho aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT leedongki aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT kimsoyoun aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis AT jeongokchan aptameraffinitybeadmediatedcaptureanddisplacementofgramnegativebacteriausingacoustophoresis |