Citation: XIONG Meng, REN Jin-Zhi, HU Shan-Wen, XU Jing-Juan, CHEN Hong-Yuan. Integrated Microfluidic Chip for Aptamer Affinity Probe Electrophoresis Assay Using in Situ Photopatterned Nanoporous Membrane[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(4): 602-610. doi: 10.19756/j.issn.0253-3820.201694 shu

Integrated Microfluidic Chip for Aptamer Affinity Probe Electrophoresis Assay Using in Situ Photopatterned Nanoporous Membrane

1.
School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212018, China;
2.
School of Public Health, Fujian Medical University, Fuzhou 350122, China;
3.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China

Corresponding author: XU Jing-Juan, xujj@nju.edu.cn
Received Date: 21 November 2020
Revised Date: 4 February 2021

Fund Project: Supported by the National Natural Science Foundation of China (Nos.21705059, 21804064).

Herein we proposed an integrated microfluidic chip to perform an aptamer-based affinity electrophoresis assay for the analysis of trace-level protein. The integrated platform included two functional units, a nanoporous membrane for sample preconcentration to improve detection sensitivity and a long gel sieving matrix for effective sample separation. The spatially controlled photopatterning technology was used to create poly(N-isopropylacrylamide) (PNIPAAm) nanoporous membrane microstructure inside the glass microchip, and methylcellulose (MC) gel solution was perfused adjacent to the membrane in the separation channel enabled zero dead volume. A model aptamer targeting thrombin was demonstrated to evaluate the performance of integrated microchip, and a simple double-T microchip was designed as a reference without the preconcentration unit. A confocal laser-induced fluorescence detection system (LIF) was utilized to quantitate the target protein thrombin. After 30 s of preconcentration and 120 s of separation, the integrated microfluidic chip showed significant signal enhancement and good separation resolution of thrombin complex with a detection range of 1-40 nmol/L and a detection limit of 1 nmol/L (S/N=3), which was over 100-fold lower than a simple double-T microchip (the detection limit was 100 nmol/L, S/N=3). The integrated platform presented in this work is anticipated to improve sensitivity of other affinity electrophoresis-based analytes.
- Photopatterning,
- Poly(N-isopropylacrylamide),
- Preconcentration,
- Integrated microfluidic chip,
- Aptamer affinity electrophoresis
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