Protein characterization

From PTM Profiling to Full Protein Characterization

The pattern of modifications on a protein can have significant influence on its functionality, activity level,  solubility, or it’s half-life. The ability to accurately detect, profile and characterize post translational modifications (PTMs) on proteins in a sample is essential to the understanding of biological systems.

Comprehensive protein and PTMs characterization raise several challenges, including, maximizing sequence coverage, detecting low abundant modified peptides and dealing with the fragmentation challenges encountered by specific modifications like glycosylation or phosphorylation.

Maximized sequence coverage

Maximizing sequence coverage is a must for applications ranging from protein product production control to the mapping of PTMs in a shotgun protoemics experiment. The combination of the additional ion mobility separation and of the timsTOF Pro’s high quality MS/MS spectra @ > 100 Hz is key to maximizing the number of fragmented peptides and hence the sequence coverage of a protein. For antibody or biosimilar drug production control, for instance, 100% sequence coverage of both the heavy and hight chain can be obtained from a very short 10 min gradient using the timsTOF Pro.

Detecting more PTM’s

Maximizing sequence coverage is a must for applications ranging from protein product production control to the mapping of PTMs in a shotgun Proteomics experiment.

PASEF
Powered by PASEF
PASEF allows to separate and fragment
PASEF allows to separate and fragmentation @>100Hz two coeluted peptides which m/z differs only of 0,9 mDa. Without separation from ion mobility dimension, the fragmentation of these two peptides would have resulted in a chimeric MS/MS spectrum.

Maximizing MS/MS information

Unlike a resonant excitation CID, CID in the quadrupolar collision cell produces a more energetic and comprehensive fragmentation pattern. Peptide sequence information can be directly obtained from phosphopeptides MS2 spectra. The timsTOF PRO also features the unique ability to acquire different collision energies for a given precursor,  producing optimal fragmentation of both the glycan and peptide moiety when analyzing glycopeptides.

For Research Use Only. Not for Use in Clinical Diagnostic Procedures.