Integrated microfluidic system for rapid screening of CRP aptamers utilizing systematic evolution of ligands by exponential enrichment (SELEX).
The systematic evolution of ligands by exponential enrichment (SELEX) is an experimental procedure that allows screening of given molecular targets by desired binding affinities from an initial random pool of oligonucleotides and oligomers. The final products of SELEX are usually referred as aptamers, which are recognized as promising molecules for a variety of biomedical applications. However, SELEX is an iterative process requiring multiple rounds of extraction and amplification that demands significant time and labor. Therefore, this study presents a novel, automatic, miniature SELEX platform. As a demonstration, the rapid screening of C-reactive protein (CRP) aptamers was performed. By utilizing microfluidic technologies and magnetic beads conjugated with CRP, aptamers with a high affinity to CRP were extracted from a random single-strand deoxyribonucleic acid (ssDNA) pool. These aptamers were further amplified by an on-chip polymerase chain reaction (PCR) process. After five consecutive extraction and amplification cycles, a specific aptamer with the highest affinity was screened automatically. The screened aptamers were used as a recognition molecule for the detection of CRP. The developed microsystem demonstrated fast screening of CRP aptamers and can be used as a powerful tool to select analyte-specific aptamers for biomedical applications.
Huang
Chao-June
CJ
Department of Engineering Science, National Cheng Kung University, 1, University Road, Tainan, Taiwan.
Lin
Hsin-I
HI
Shiesh
Shu-Chu
SC
Lee
Gwo-Bin
GB
D052157:Aptamers, Nucleotide / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002097:C-Reactive Protein / Q000032:analysis* / Q000235:genetics*; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008024:Ligands; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results; D052156:SELEX Aptamer Technique / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D018511:Systems Integration
D052157:Aptamers, Nucleotide; D008024:Ligands; D002097:C-Reactive Protein