Herein, we present a large-area 3D hemispherical perforated microwell structure for a bead based bioassay. Such a unique microstructure enables us to perform the rapid and stable localization of the beads at the single bead level and the facile manipulation of the bead capture and retrieval with high speed and efficiency. The fabrication process mainly consisted of three steps: the convex micropatterned nickel (Ni) mold production from the concave micropatterned silicon (Si) wafer, hot embossing on the polymer matrix to generate the concave micropattened acrylate sheet, and reactive ion etching to make the bottom holes. The large-area hemispherical perforated micropatterned acrylate sheet was sandwiched between two polydimethylsiloxane (PDMS) microchannel layers. The bead solution was injected and recovered in the top PDMS microchannel, while the bottom PDMS microchannel was connected with control lines to exert the hydrodynamic force in order to alter the flow direction of the bead solution for the bead capture and release operation. The streptavidin-coated microbead capture was achieved with almost 100% yield within 1 min, and all the beads were retrieved in 10 s. Lysozyme or thrombin binding aptamer labelled microbeads were trapped on the proposed bead microarray, and the in situ fluorescence signal of the bead array was monitored after aptamer-target protein interaction. The protein-aptamer conjugated microbeads were recovered, and the aptamer was isolated for matrix assisted laser desorption/ionization time-of-flight mass spectrometry analysis to confirm the identity of the aptamer.
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November 2014
Research Article|
December 09 2014
A large-area hemispherical perforated bead microarray for monitoring bead based aptamer and target protein interaction
Jong Seob Choi;
Jong Seob Choi
Department of Chemical and Biomolecular Engineering (BK21 program) and Institute for the BioCentury,
Korea Advanced Institute of Science and Technology (KAIST)
, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, South Korea
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Sunwoong Bae;
Sunwoong Bae
Department of Chemical and Biomolecular Engineering (BK21 program) and Institute for the BioCentury,
Korea Advanced Institute of Science and Technology (KAIST)
, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, South Korea
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Kyung Hoon Kim;
Kyung Hoon Kim
Department of Chemical and Biomolecular Engineering (BK21 program) and Institute for the BioCentury,
Korea Advanced Institute of Science and Technology (KAIST)
, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, South Korea
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Tae Seok Seo
Tae Seok Seo
a)
Department of Chemical and Biomolecular Engineering (BK21 program) and Institute for the BioCentury,
Korea Advanced Institute of Science and Technology (KAIST)
, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, South Korea
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a)
Author to whom correspondence should be addressed. Electronic mail: [email protected]
Biomicrofluidics 8, 064119 (2014)
Article history
Received:
October 24 2014
Accepted:
November 26 2014
Citation
Jong Seob Choi, Sunwoong Bae, Kyung Hoon Kim, Tae Seok Seo; A large-area hemispherical perforated bead microarray for monitoring bead based aptamer and target protein interaction. Biomicrofluidics 1 November 2014; 8 (6): 064119. https://doi.org/10.1063/1.4903939
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