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Monitoring of enzymatic cleavage reaction of GST-fusion protein on biolayer interferometry sensor.
Biophys Physicobiol
September 2024
Faculty of Pharmaceutical Sciences, Hokkaido University of Science, Sapporo, Hokkaido 006-8585, Japan.
Biolayer interferometry (BLI) is an optical sensor-based analytical method primarily used for analyzing interactions between biomolecules. In this study, we explored the application of BLI to observe the cleavage reaction of glutathione S-transferase (GST)-tagged fusion protein by human rhinovirus (HRV) 3C protease on a BLI sensor as a new application of the BLI method. The soluble domain of the Tic22 protein from was expressed and purified as a GST-tagged fusion protein, GST-Tic22, in .
View Article and Find Full Text PDFMagnetic relaxation switch biosensor for detection of Vibrio parahaemolyticus based on photocleavable hydrogel.
Anal Chim Acta
January 2025
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, PR China. Electronic address:
Background: Foodborne pathogens, particularly Vibrio parahaemolyticus (VP) found in seafood, pose significant health risks, including abdominal pain, nausea, and even death. Rapid, accurate, and sensitive detection of these pathogens is crucial for food safety and public health. However, existing detection methods often require complex sample pretreatment, which limits their practical application.
View Article and Find Full Text PDFGold nanocube-enhanced SERS biosensor based on heated electrode coupled with exonuclease III-assisted cycle amplification for sensitive detection of flap endonuclease 1 activity.
Talanta
December 2024
Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian, 350108, China. Electronic address:
The flap endonuclease 1 (FEN1) plays a key role in DNA replication and repair, its aberrant expression is associated with tumor development, so it has been recognized as a promising biomarker for a variety of cancers. Here, a novel "turn on" mode gold nanocube-enhanced surface-enhanced Raman scattering (SERS) biosensor was constructed by combining a heated Au electrode (HAuE), exonuclease III (Exo III)-assisted cycle amplification, and gold nanocube (AuNC)-based SERS enhancement to achieve highly sensitive detection of FEN1 activity. The SERS tag was prepared using the Raman reporter modified on the AuNC surface, and the high electromagnetic field provided by the sharp geometric feature of AuNC greatly enhanced the SERS signal.
View Article and Find Full Text PDFProduction of virus-like particles of FMDV by 3C protease cleaving precursor polyprotein P1 in vitro.
Appl Microbiol Biotechnol
December 2024
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730000, China.
Nonstructural protein 3C, a master protease of Picornaviridae, plays a critical role in viral replication by directly cleaving the viral precursor polyprotein to form the viral capsid protein and antagonizing the host antiviral response. Additionally, 3C protease, as a tool enzyme, is involved in regulating polyprotein expression. Here, the 3C mutant gene (3Cm), fused with a small ubiquitin-like modifier (SUMO) tag at the N-terminal and featuring a mutation at position 127, was inserted into the cold-shock plasmid pCold of Escherichia coli for expression.
View Article and Find Full Text PDFImprovements in large-scale production of tobacco etch virus protease.
Protein Expr Purif
December 2024
Protein Expression Laboratory, NCI RAS Initiative, Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, USA.
Tobacco-etch-virus (TEV) protease is the workhorse of many laboratories in which protein expression is the linchpin of downstream experiments. TEV protease is remarkable in its sequence specificity as the cleavage sequence rarely appears in higher organisms and its ability to cleave fusion tag proteins from proteins of interest. Herein we report work done on large-scale production of TEV protease using different promotors, media, fusion tags, and expression platforms.
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