September 19 - 22, 2017
Boston, MA, USA
The Genedata team of experts will be at booth #1 at the 14th Symposium on the Practical Applications of Mass Spectrometry in the Biotechnology Industry to demonstrate all the new features of Genedata Expressionist.
To schedule a meeting in advance, please contact Genedata at email@example.com.
An adapted top-down LC-MS approach for screening lipid-modified proteins
Andrew James, Sanofi Pasteur, Toronto, ON, Canada
Technical Lunch Seminar - Beyond Oncology Session
Thursday, September 21 | 12:15 - 12:45
In vaccine development, liquid chromatography-mass spectrometry (LC-MS) is typically applied to characterization and identity workflows, particularly for recombinant protein candidates. Intact mass analysis by MS is particularly useful for establishing product identity, for both therapeutic proteins and recombinant vaccine antigens. However, when the protein of interest undergoes post-translational modification (PTM) during expression, intact mass analysis of the heterogeneous product is more complex, and confirming identity becomes difficult. Often, alternate MS workflows, such as a peptide mapping, are required to establish identity.
This work focuses on the development of an intact mass LC-MS method for the analysis of lipid modified proteins. The method was originally evaluated for detection of the different protein ‘lipoforms’. By applying in-source fragmentation within the workflow, we were able to determine protein sequence identity and further characterize the lipid profiles of individual lipoprotein populations.
Lipid modifications are made on the N-terminal Cysteine of selected proteins during bacterial expression. This creates a heterogeneously modified Cys residue that is more susceptible to in-source fragmentation during MS experiments. The rest of protein remains intact, and can be identified in a sequence specific manner using mass deconvolution. Meanwhile, the lipidated Cys fragment can be detected and further analyzed to characterize and confirm specific PTM structures. As this method evolves, we will apply principals of multiple attribute monitoring to design an assay with readouts for protein identity, detection of protein lipoforms, lipid identification, and protein purity.
Automated Data Processing for Quality Monitoring of Biotherapeutics by Multi-attribute Methods (MAMs)
Maurizio Bronzetti, Genedata
Automated Intact Mass Analysis for the Characterization of Antibodies
Peter Haberl, Genedata
Automated Workflow for Host Cell Protein Monitoring by Mass Spectrometry: From Raw Data to Final Report
Joe Shambaugh, Genedata
Enterprise Mass Spectrometry Software Solution Enabling Characterization of Biotherapeutics from Discovery and Development to Production and Quality Control
Cassandra Wigmore, Genedata
Monitoring critical quality attributes of an IgG4 antibody during process development with Genedata Expressionist
Larry Wang, Merck