Title :
Isolation of thermally sensitive aptamers on a microchip
Author :
Hilton, J.P. ; Kim, Jinho ; Nguyen, ThaiHuu ; Barbu, Mihaela ; Pei, Renjun ; Stojanovic, Milan ; Lin, Qiao
Author_Institution :
Dept. of Mech. Eng., Columbia Univ., New York, NY, USA
fDate :
Jan. 29 2012-Feb. 2 2012
Abstract :
We present a microchip for isolation of aptamers that bind to target ligands at prespecified temperatures. The device uses integrated resistive heaters and sensors to control the temperatures of (poly)dimethylsiloxane (PDMS) microchambers for temperature-specific selection and bead-based amplification of aptamers. Aptamers are isolated from a randomized DNA library at specified temperatures, and amplified onto microbeads using bead-based polymerase chain reaction (PCR). As a proof of concept, the device was used to isolate aptamers for human immunoglobulin E (IgE), with the enriched pool of candidate aptamers exhibiting a much higher and temperature-dependent affinity for the target protein. The procedure was performed with significantly less reagent use in a much shorter time period (4 hours) than with conventional devices (up to 2 days).
Keywords :
DNA; biochemistry; biosensors; molecular biophysics; polymers; proteins; (poly)dimethylsiloxane microchambers; aptamer isolation; bead-based amplification; bead-based polymerase chain reaction; human immunoglobulin E; integrated resistive heaters; integrated resistive sensors; microchip; protein; randomized DNA library; temperature-specific selection; thermally sensitive aptamers; time 4 hour; Artificial neural networks; DNA; Fluorescence; Libraries; Microfluidics; System-on-a-chip; Temperature sensors;
Conference_Titel :
Micro Electro Mechanical Systems (MEMS), 2012 IEEE 25th International Conference on
Conference_Location :
Paris
Print_ISBN :
978-1-4673-0324-8
DOI :
10.1109/MEMSYS.2012.6170103