27. Development of a Synthetic Protein Quality Control (QC) Standard for the Assessment of Sample Proteolysis Reproducibility
Mon 12:18 PM - PosterSplash Track 3
Bryan Krastins
ThermoFisher Scientific
View Long Abstract
Because enzymatic digestion is the starting point for many proteomic experiments and assays, inconsistent sample preparation can compromise the downstream analysis of the resulting data, especially in SRM assays where accurate quantitation of peptides is crucial. Therefore, a thorough understanding of proteolytic digestion efficiency during the sample preparation process is critical to the success of these methods. A synthetic protein which could be used to assess the quality and efficiency of proteolytic digestion during sample preparation was developed. The proteolytic QC protein was tested by adding a known concentration to a crude serum sample and then subjecting the mixture to enzymatic digestion followed by mass spectrometry analysis in a triple quadrupole. This approach enabled quantification of proteins from the serum sample with simultaneous assessment of proteolytic efficiency.