Improving aptamer selection efficiency through volume dilution, magnetic concentration, and continuous washing in microfluidic channels
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- Title
- Improving aptamer selection efficiency through volume dilution, magnetic concentration, and continuous washing in microfluidic channels
- Authors
- Oh, SS; Ahmad, KM; Cho, M; Kim, S; Xiao, Y; Soh, HT
- Date Issued
- 2011-09-01
- Publisher
- American Chemical Society
- Abstract
- The generation of nucleic acid aptamers with high affinity typically entails a time-consuming, iterative process of binding, separation, and amplification. It would therefore be beneficial to develop an efficient selection strategy that can generate these high-quality aptamers rapidly, economically, and reproducibly. Toward this goal, we have developed a method that efficiently generates DNA aptamers with slow off-rates. This methodology, called VDC-MSELEX, pairs the volume dilution challenge process with microfluidic separation for magnetic bead-assisted aptamer selection. This method offers improved aptamer selection efficiencies through the application of highly stringent selection conditions: it retrieves a small number (<10(6)) of magnetic beads suspended in a large volume (>50 mL) and concentrates them into a microfluidic chamber (8 mu L) with minimal loss for continuous washing. We performed three rounds of the VDC-MSELEX using streptavidin (SA) as the target and obtained new DNA aptamer sequences with low nanomolar affinity that specifically bind to the SA proteins.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/37726
- DOI
- 10.1021/AC201269F
- ISSN
- 0003-2700
- Article Type
- Article
- Citation
- ANALYTICAL CHEMISTRY, vol. 83, no. 17, page. 6883 - 6889, 2011-09-01
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