Low-Artifact Digestion Buffer (EMS0011) is optimized for digestion of therapeutic proteins and antibodies prior to peptide mapping by LC-UV-MS. The buffer has been designed to minimize artifactual methionine oxidation and asparagine deamidation during sample preparation while maintaining effective reduction, alkylation and proteolytic digestion.
Endogenous deamidation and oxidation of protein amino acids can be implicated in and be indicative of many diseases, protein turnover, development, and aging. The introduction of these modifications can often affect biological activity, half-life, and immunogenicity.
Current protein digestion workflows often introduce a significant amount of artifactual deamidation and oxidation which prohibits the measurement of the endogenous levels.
The Low Artifact Digestion Buffer allows protein digestion in less than 6 hours with minimized artifactual modifications such as deamidation and oxidation. The amount of oxidation and deamidation introduced during further downstream processing of biotherapeutic antibodies and proteins can be accurately determined. The most commonly deamidated amino acid is Asn especially when followed by small amino acids such as Gly. The most commonly oxidized amino acids are Met, Cys, and to some extent Trp and His.