Assembly of Protein Nanopore in Microfluidic Tube and Its Application in Single-Molecule Analysis
10.19756/j.issn.0253-3820.251089
- VernacularTitle:微流管蛋白纳米孔组装及单分子分析
- Author:
Peng ZHENG
1
;
Xin-Yu SHI
;
Xing-Peng WEI
;
Xiao-Feng KANG
Author Information
1. 山东飞扬化工有限公司,泰安 271200
- Keywords:
Protein nanopore;
Microfluidic system;
Single-molecule analysis;
Lipid bilayer;
Microprocessing
- From:
Chinese Journal of Analytical Chemistry
2025;53(7):1090-1096
- CountryChina
- Language:Chinese
-
Abstract:
As the simplest label-free single-molecule analytical technique in solution,nanopore technology has been widely used in rapid sequencing of protein and nucleic acid,and single-molecule sensing.However,the main challenge is to get this technology out of the lab and achieve automation,miniaturization as well as rapid analysis at high throughput.In this work,a new microfluidic tubing protein nanopore analysis system was designed and constructed by combining microfluidic and nanopore nanofluidic analysis techniques.An arcuate lipid bilayer was formed in the microfluidic device through lipid monolayer at the oil/water interface by developed aperture processing technology at 10 μm level.The arcuate lipid bilayer with the life span of more than 98 h had high biomimetic fluidity and biological activity by which protein channel could be produced and sealed at 18 GΩ level without leakage.When applied+40 mV potential,relative standard deviation of single proteinα-hemolysin mutant(α-HL(M113F/K147N))channel current was less than 0.16 pA(n=3)in 10 mmol/L Tris-HCl(pH 7.4)buffer containing 1 mol/L NaCl.Single-molecule detection for aptamer and its host-guest interactions with biotin in protein channel demonstrated that this microfluidic tube protein nanopore sensor could well work with high sensitivity without the need of anti-vibration table and shielding cage.
