Improvements in Workflow Capabilities to Provide Increased Reproducibility, Speed and Throughput for Quantitation and Characterisation in Peptide Mapping of Biotherapeutics

Robert van Ling, European Expert Chromatography Columns & Consumables, Thermo Fisher Scientific, BREDA, Netherlands
Mike Oliver, Thermo Scientific, Runcorn, United Kingdom

Protein digestion is a fundamental technique employed in biopharmaceutical applications, often taking advantage of proteomics digestion protocols. Despite its widespread use, protein digestion may still provides many analytical challenges. The optimum digestion requires conditions that:
• Cleave the proteins after every lysine and arginine
• Unfold the protein/proteins of interest
• Reduces autolysis effects

This can be a lengthy and complex process and often leads to poor reproducibility.

In this presention, we describe a new and simple digestion platform for biotherapeutics. This novel workflow removes uncertainties associated with conventional solution-based tryptic digestion protocols. Applying immobilized and heat-stable trypsin technology, combined with the optimized buffer conditions, this results in higher reproducibility and superior sample characterization.