New Workflow for Top-Down Protein Sequence Confirmation and de novo Sequencing
Bruker Scientific, "New Workflow for Top-Down Protein Sequence Confirmation and de novo Sequencing," with Mariangela Kosmopoulou, Analytical Chemist, and Arndt Asperger, Senior Application Scientist.
This webinar will demonstrate how to apply innovative top-down algorithms to:
- Get fast sequencing results with confidently annotated top-down MS spectra
- Identify unknown proteins from top-down MS data
- Rapidly confirm protein terminal sequences with a complete workflow based on the timsTOF fleX system
Primary sequence determination and characterization of heterogeneities is an essential task for protein development. Top-down analysis can support these by sequencing N and C- terminals even if modified.
The mass spectral complexity encountered in high resolution/accurate mass spectrometry data previously limited Top-Down analyses to sequence confirmation. Thus, the de novo determination of protein sequences without pre-existing sequence information requires new data analysis software to reliably extract and timely process such high-density information. Here, we report on the development of new workflows for protein sequence confirmation and de novo sequencing by processing complex Top-Down MS/MS data with the Bruker OmniScape software.
Amongst top-down MS/MS methods, MALDI Top-Down Sequencing (MALDI-TDS) stands out in that it is a fragment-all method relying on In-Source Decay (ISD) producing fragment ions of low charge states (singly charged mainly) simplifying data analysis. When performing MALDI-TDS on a timsTOFfleX instrument, Trapped Ion mobility Spectrometry (TIMS) can serve as an added dimension of fragment ion separation regarding charge state and terminal origin. OmniScape software enables analysis of data obtained from TIMS enabled MALDI-TDS analyses in a simple and fast workflow providing confident sequence verification of a target protein including its terminal status with shortest time to result.
Mariangela Kosmopoulou is an analytical scientist at Bruker with background on pharmaceutical chemistry and mass spectrometry expertise including sample prep, LC-MS method development, data analysis and biochemical elucidation of results. Kosmopoulou is currently guiding software development, testing and evaluating beta versions for top-down analysis of proteins and conducting software documentation and trainings.
Arndt Asperger, PhD, is senior application scientist at Bruker Daltonics GmbH & Co. KG, Bremen, Germany. Arndt is analytical chemist by training and got his PhD from Leipzig University. He joined Bruker in 2002 and since then has been working in various application related positions with a focus on pharma/biopharma workflows.