Webinar : Enzymatic Workflows for Protein Characterization

This webinar presents an integrated enzymatic workflow for comprehensive structural characterization of therapeutic proteins, including monoclonal antibodies, biosimilars, and recombinant proteins. The presentation demonstrates how PNGase F, Trypsin MS Grade, and Carboxypeptidase B function synergistically to enable complete protein analysis through N-glycan profiling, peptide mapping with post-translational modification (PTM) identification, and C-terminal characterization.

In the development and quality control of biotherapeutics, detailed characterization of critical quality attributes (CQAs) is essential for ensuring safety, efficacy, and batch-to-batch consistency. N-glycosylation profoundly influences pharmacokinetics, effector functions, and potential immunogenicity, necessitating precise profiling of glycan heterogeneity using PNGase F for enzymatic release and subsequent analysis. Peptide mapping, facilitated by high-grade trypsin digestion, remains a cornerstone technique for confirming primary amino acid sequence integrity, detecting site-specific PTMs (such as oxidation, deamidation, and glycosylation), and identifying sequence variants or truncations—key requirements in regulatory submissions for both novel therapeutics and biosimilars. Complementing these, Carboxypeptidase B treatment enables accurate assessment of C-terminal heterogeneity, including lysine clipping, which affects charge profiles and product-related impurities often monitored via ion-exchange chromatography or mass spectrometry.

This multi-enzyme approach addresses limitations of single-method analyses by providing orthogonal, synergistic data that support robust comparability exercises, process optimization, and compliance with stringent regulatory expectations (e.g., ICH Q6B guidelines and biosimilar similarity assessments). Attendees will gain insight into the scientific rationale underpinning this workflow, interpret analytical outputs from each enzymatic step, and understand its practical application in resolving CQAs critical for successful regulatory filings and biopharmaceutical development.

Speaker

Dr. Pavan Kumar Avula, PhD
Senior Manager – Techno Commercial, Provis Biolabs

Dr. Avula holds a PhD in Neuroscience from the University of Melbourne and a Master's degree in Biotechnology from La Trobe University. As Senior Manager at Provis Biolabs, he specializes in supporting analytical workflows for therapeutic protein characterization, working closely with biopharmaceutical researchers and QC laboratories. His role combines scientific understanding with practical implementation expertise.