Microchip CE/MS Peptide Mapping Workflow with Trypsin Digestion for the Analysis of Critical Quality Attributes of Monoclonal Antibodies

By Sara Carillo, Rachel Ronan, Silvia Millán-Martín, Adi Kulkarni, Erin Redman, Kate Yu, Jonathan Bones

A new study has optimized a peptide mapping protocol that's compatible with ZipChip CE-MS analysis, significantly reducing the time required for analysis. The method uses a Thermo Scientific Orbitrap Exploris 240 Mass Spectrometer and can complete the entire peptide mapping analysis workflow in less than two hours. It offers 99% sequence coverage and accurate determination of critical quality attributes, making it a faster and more efficient alternative to traditional LC-based approaches.

The method is especially useful for high-throughput analysis and evaluation of monoclonal antibodies. The study also developed an optimized protocol for sample preparation compatible with ZipChip CE-MS analysis. The results showed no significant differences between samples prepared with the modified protocol and those prepared with the standard protocol. The protocol ensures high sample reproducibility and low artificially induced modifications, making it suitable for use in a Multi-Attribute Method (MAM) workflow. The ZipChip CE-MS preparation workflow offers quicker analysis times, making it a faster alternative to traditional LC-based approaches. This method can be easily implemented in both academic and industrial characterization laboratories.