Acousto-microfluidics for screening of ssDNA aptamer

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Acousto-microfluidics for screening of ssDNA aptamer. / Park, Jeewoong; Lee, Sujin; Ren, Shuo; Lee, Sangwook; Kim, Soyoun; Laurell, Thomas.

In: Scientific Reports, Vol. 6, 27121, 08.06.2016.

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Park, Jeewoong ; Lee, Sujin ; Ren, Shuo ; Lee, Sangwook ; Kim, Soyoun ; Laurell, Thomas. / Acousto-microfluidics for screening of ssDNA aptamer. In: Scientific Reports. 2016 ; Vol. 6.

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TY - JOUR

T1 - Acousto-microfluidics for screening of ssDNA aptamer

AU - Park, Jeewoong

AU - Lee, Sujin

AU - Ren, Shuo

AU - Lee, Sangwook

AU - Kim, Soyoun

AU - Laurell, Thomas

PY - 2016/6/8

Y1 - 2016/6/8

N2 - We demonstrate a new screening method for obtaining a prostate-specific antigen (PSA) binding aptamer based on an acoustofluidic separation (acoustophoreis) technique. Since acoustophoresis provides simultaneous washing and separation in a continuous flow mode, we efficiently obtained a PSA binding aptamer that shows high affinity without any additional washing step, which is necessary in other screening methods. In addition, next-generation sequencing (NGS) was applied to accelerate the identification of the screened ssDNA pool, improving the selecting process of the aptamer candidate based on the frequency ranking of the sequences. After the 8 th round of the acoustophoretic systematic evolution of ligands by exponential enrichment (SELEX) and following sequence analysis with NGS, 7 PSA binding ssDNA aptamer-candidates were obtained and characterized with surface plasmon resonance (SPR) for affinity and specificity. As a result of the new SELEX method with PSA as the model target protein, the best PSA binding aptamer showed specific binding to PSA with a dissociation constant (K d) of 0.7 nM.

AB - We demonstrate a new screening method for obtaining a prostate-specific antigen (PSA) binding aptamer based on an acoustofluidic separation (acoustophoreis) technique. Since acoustophoresis provides simultaneous washing and separation in a continuous flow mode, we efficiently obtained a PSA binding aptamer that shows high affinity without any additional washing step, which is necessary in other screening methods. In addition, next-generation sequencing (NGS) was applied to accelerate the identification of the screened ssDNA pool, improving the selecting process of the aptamer candidate based on the frequency ranking of the sequences. After the 8 th round of the acoustophoretic systematic evolution of ligands by exponential enrichment (SELEX) and following sequence analysis with NGS, 7 PSA binding ssDNA aptamer-candidates were obtained and characterized with surface plasmon resonance (SPR) for affinity and specificity. As a result of the new SELEX method with PSA as the model target protein, the best PSA binding aptamer showed specific binding to PSA with a dissociation constant (K d) of 0.7 nM.

UR - http://www.scopus.com/inward/record.url?scp=84976448852&partnerID=8YFLogxK

U2 - 10.1038/srep27121

DO - 10.1038/srep27121

M3 - Article

VL - 6

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 27121

ER -