Clean and Complete Protein Digestion with an Autolysis Resistant Trypsin for Peptide Mapping.

ABSTRACT: Peptide mapping requires cleavage of proteins in a predictable fashion so that target protein-specific peptides can be reliably identified and quantified. Trypsin, a commonly used protease in this process, can also undergo self-cleavage or autolysis, thereby reducing the effectivity and even cleavage specificity at lysine and arginine residues. Here, we report highly efficient and reproducible peptide mapping of biotherapeutic monoclonal antibodies. We highlight the properties of a homogeneous chemically modified trypsin on thermal stability, a 54% increase in melting temperature with an 84% increase in energy required for unfolding, an indication of more thermally stable trypsin, >90% retained intact mass peak area after exposure to digestion conditions confirming autolysis resistance, 10

SUBMITTER: Muriithi B 

PROVIDER: S-EPMC11536465 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature