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An automatic microfluidic system that continuously performs the systematic evolution of ligands by exponential enrichment

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Abstract

The systematic evolution of ligands by exponential enrichment (SELEX) technique has been extensively used to screen molecule-specific aptamers from combinatorial libraries of synthetic nucleic acids. Aptamers are single-stranded DNA or RNA, which have a high affinity to a large variety of molecules ranging from small drugs or metabolites to cells. Therefore, they have a variety of promising applications such as for diagnostics and targeted therapeutics. In this study, a new microfluidic chip was developed to perform continuous screening of DNA-based aptamers in an automatic format. When compared with the existing manual procedure, the developed microfluidic chip has several advantages including a rapid and efficient screening process, automation, and less consumption of samples/reagents. Experimental data showed that an aptamer specific to alpha-fetoprotein was successfully screened from a random DNA pool. The entire screening process (five continuous, repetitive rounds) can be completed within 6 h, which is much faster than the traditional methods (more than 15 h). An automatic, rapid and efficient SELEX process was performed by this developed microfluidic chip, which may enable a generalized platform for the fast screening of DNA-based biomarkers in the future.

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Abbreviations

AFP:

Alpha-fetoprotein

ASIC:

Application-specific integrated circuit

BSA:

Bovine serum albumin

CCD:

Charge-coupled device

CNC:

Computer-numerical-control

DI:

Deionized

EMVs:

Electromagnetic valves

MEMS:

Micro-electro-mechanical-systems

PBST:

Phosphate buffer saline with 1 % of Tween 20

PCR:

Polymerase chain reaction

PDMS:

Polydimethylsiloxane

RLU:

Relative light unit

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SELEX:

Systematic evolution of ligands by exponential enrichment

SEM:

Scanning electron microscope

ssDNA:

Single-stranded deoxyribonucleic acid

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Acknowledgments

The authors would like to thank the National Science Council in Taiwan (NSC 100-2120-M-007-014) and the “Toward A World-class University” Project for financial support of this study.

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Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, Tainan, 701, Taiwan

    Yin-He Chen & Chao-Jyun Huang

  2. Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, Tainan, 701, Taiwan

    Hsin-I Lin & Shu-Chu Shiesh

  3. Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, 30013, Taiwan

    Gwo-Bin Lee

Authors
  1. Yin-He Chen
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  2. Hsin-I Lin
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  3. Chao-Jyun Huang
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  4. Shu-Chu Shiesh
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  5. Gwo-Bin Lee
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Corresponding authors

Correspondence to Shu-Chu Shiesh or Gwo-Bin Lee.

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Chen, YH., Lin, HI., Huang, CJ. et al. An automatic microfluidic system that continuously performs the systematic evolution of ligands by exponential enrichment. Microfluid Nanofluid 13, 929–939 (2012). https://doi.org/10.1007/s10404-012-1013-8

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  • DOI: https://doi.org/10.1007/s10404-012-1013-8

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