2. Academic Validation
  3. A novel assay for the determination of PreScission protease by capillary electrophoresis

A novel assay for the determination of PreScission protease by capillary electrophoresis

  • Biophys Chem. 2022 Feb:281:106696. doi: 10.1016/j.bpc.2021.106696.
Shuwen Zhou 1,  Pengfei Cui 1,  Jingyu Sheng 2,  Xueli Zhang 3,  Pengju Jiang 4,  Xinye Ni 5,  Kai Cao 6,  Lin Qiu 7
Affiliations

Affiliations

  • 1 School of Pharmacy, Changzhou University, Changzhou 213164, Jiangsu, PR China; The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213003, PR China.
  • 2 Wujin Hospital Affiliated with Jiangsu University, Changzhou 213017, Jiangsu, PR China; The Wujin Clinical College of Xuzhou Medical University, Changzhou 213017, Jiangsu, PR China.
  • 3 Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, PR China.
  • 4 School of Pharmacy, Changzhou University, Changzhou 213164, Jiangsu, PR China.
  • 5 The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213003, PR China. Electronic address: nxy@njmu.edu.cn.
  • 6 Wujin Hospital Affiliated with Jiangsu University, Changzhou 213017, Jiangsu, PR China; The Wujin Clinical College of Xuzhou Medical University, Changzhou 213017, Jiangsu, PR China. Electronic address: caokai9001@126.com.
  • 7 School of Pharmacy, Changzhou University, Changzhou 213164, Jiangsu, PR China. Electronic address: linqiupjj@gmail.com.
PMID: 34954553 DOI: 10.1016/j.bpc.2021.106696
Abstract

The detection of protease activity in the body plays a significant role in the early diagnosis of diseases. However, Enzymes inevitably come into contact with various complex biological fluids in the body during the flow, which greatly increases the detection difficulty. Therefore, protease detection in vivo has great challenges. Herein, we report a new assay for detecting protease using capillary electrophoresis inside a capillary with semicircular bends. We first designed a peptide substrate, and then the peptide was self-assembled with quantum dots to form a QDs-peptide substrate. The capillary was bent to semicircular-shaped turns and served as a micro-reactor to allow protease and substrate react in it. Due to the different electrophoretic velocity, the protease and the substrate were mixed inside the bent capillary with sequential injections and the cleavage of the substrate can be detected using capillary electrophoresis combined with Förster resonance energy transfer technology. This novel assay will greatly expand the detection of Enzyme activity in vivo.

Keywords

Capillary electrophoresis; Fluorescence; Förster resonance energy transfer; PreScission protease; Quantum dots.

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