This presentation covers the challenges and methodologies in peptide mapping of complex bispecific antibody formats. The presentation introduces a multi-enzyme approach as a new experimental setup for sequence variant analysis, emphasizing the importance of complete protein coverage to ensure the absence of critical sequence variants and modifications. The speakers, Eva and Marco, explain their roles and the collaboration between their departments at Roche Innovation Centre, Munich, detailing how advancements in lab automation and software tools have significantly increased efficiency in peptide mapping tasks.

They highlight the practical application of this multi-enzyme approach, showing its advantages over traditional single-enzyme methods. By utilizing various enzymes, they achieve higher coverage and accuracy in detecting low-abundant modifications and sequence variants. They provide a detailed explanation of the quantification strategies employed, demonstrating how the use of multiple digests can better identify and quantify modifications compared to conventional methods. The presentation concludes with real-life examples and a discussion on the optimization of enzyme ratios and quantification approaches, aiming to offer a comprehensive understanding of the benefits and implementation of the multi-enzyme strategy in antibody development.