Integrating Top-Down Mass Spectrometry and Single-Molecule Sequencing for Proteoform Analysis
Summary
Given the numerous sources of protein variation, ranging from biological factors to sample preparation and instrument settings, there is a pressing need for next-generation analytical platforms capable of driving proteoform analysis with complete molecular precision.
In this webinar, Dr. Michael Caldwell, Scientific Officer at Northwestern Proteomics Center of Excellence, will explore top-down mass spectrometry (TD-MS) and Platinum®.
Platinum is a benchtop instrument that provides single amino acid resolution of single protein molecules to characterize IL-6, including pyroglutamate (pE), a post-translational modification (PTM) produced via enzymatic and spontaneous cyclization of N-terminal glutamine (Q)/glutamate (E) residues.
Find out how TD-MS enables intact protein analysis, limiting formation of N-terminal Q/E (MS1). During MS2, analysis of daughter ions reveals modifications and artifactual formation. Platinum employs dye-labeled N-terminal amino acid recognizers that reveal primary structure and variants, and aminopeptidases that expose the next amino acid for recognition until the peptide is sequenced. TD-MS and Platinum detect not only functionally relevant regions of IL-6 primary structure, but also sites prone to pE formation.
Key Learning Points/Takeaways:
- Understand how top-down mass spectrometry (TD-MS) and Next-Generation Protein Sequencing can complement each other for comprehensive protein analysis.
- Explore methods to bridge discovery and targeted protein analysis across different instruments and detection methods.
- Learn how TD-MS distinguishes between native and artifactual proteoforms.
Speakers: Michael Caldwell, PhD – Northwestern University, Meredith Carpenter, PhD – QSI and Kenneth Skinner, PhD – QSI
Date: September 30, 2024