Year: 2024
Type: article
Abstract: High Resolution Image Download MS PowerPoint Slide Reliable enzymatic digestion underscores successful mass-spectrometry-based proteomics experiments. In this study, we compare the use of the arginine-specific protease, GingisREX, against a more traditional approach in the identification of Escherichia coli proteins. An increased number of protein identifications were noted when GingisREX was used compared to a trypsin/lys-C mixture. This improvement was attributed to the generation of fewer peptides per protein, resulting in a simpler peptide mixture. Furthermore, GingisREX exhibited increased digestion efficiency, fewer missed cleavages, and improved MS/MS data quality for higher molecular weight peptides. The data here establish GingisREX to be a protease complementary to trypsin for enhanced detection of bacterial proteins. With further optimization, GingisREX could prove to be an effective alternative to trypsin for identifying host cell proteins in biotherapeutics.
Authors: Thomas Powell, Philip Widdowson, Andreas Nägeli, Martin Ebner, Andrew Creese
Institutions: Immunocore (United Kingdom), Genovis (Sweden)
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W4403336414