Selection and elution of aptamers using nanoporous sol-gel arrays with integrated microheaters.
RNA and DNA aptamers that bind to target molecules with high specificity and affinity have been a focus of diagnostics and therapeutic research. These aptamers are obtained by SELEX (Systematic Evolution of Ligands by EXponential enrichment) often requiring more than 10 successive cycles of selection and amplification, where each cycle normally takes 2 days per cycle of SELEX. Here, we have demonstrated the use of sol-gel arrays of proteins in a microfluidic system for efficient selection of RNA aptamers against multiple target molecules. The microfluidic chip incorporates five sol-gel binding droplets, within which specific target proteins are imbedded. The droplets are patterned on top of individually addressable electrical microheaters used for selective elution of aptamers bound to target proteins in the sol-gel droplets. We demonstrate that specific aptamers bind their respective protein targets and can be selectively eluted by micro-heating. Finally, our microfluidic SELEX system greatly improved selection efficiency, reducing the number of selection cycles needed to produce high affinity aptamers. The process is readily scalable to larger arrays of sol-gel-embedded proteins. To our knowledge, this is the first demonstration of a chip-based selection of aptamers using microfluidics, thereby allowing development of a high throughput and efficient SELEX procedures.
Park
Seung-Min
SM
Applied and Engineering Physics, Cornell University, Ithaca, NY 14853, USA.
Ahn
Ji-Young
JY
Jo
Minjoung
M
Lee
Dong-Ki
DK
Lis
John T
JT
Craighead
Harold G
HG
Kim
Soyoun
S
D052157:Aptamers, Nucleotide / Q000235:genetics / Q000302:isolation & purification*; D017076:Computer-Aided Design; D004586:Electrophoresis / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006361:Heating / Q000295:instrumentation*; D008839:Microelectrodes; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008904:Miniaturization; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000295:instrumentation; D044367:Phase Transition; D016062:Porosity; D015203:Reproducibility of Results; D052156:SELEX Aptamer Technique / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D018511:Systems Integration; D014462:Ultrafiltration / Q000295:instrumentation*
D052157:Aptamers, Nucleotide