Peptide characterization
Detailed sequence analysis by quantitative peptide mapping
Getting a peptide map is a critical part of any peptide characterization project. You use it to confirm the amino acid sequence of your protein and quantify about post-translational modifications (PTMs). Peptide analysis can even locate truncations and cleavage sites, as well as identify degradation products with stability studies.
The analysis is often...
Getting a peptide map is a critical part of any peptide characterization project. You use it to confirm the amino acid sequence of your protein and quantify about post-translational modifications (PTMs). Peptide analysis can even locate truncations and cleavage sites, as well as identify degradation products with stability studies.
The analysis is often a vital part of comparability studies – especially in quality assurance/quality control (QA/QC) of protein-based biologics. These include mAbs, ADCs, and also recombinant proteins.
How to get high sequence coverage with specific proteases
The principle uses a specific protease that digests the protein and turns it into peptides. We then perform peptide characterization by LC-MS/MS. More specifically, we use the peptide mass (MS) and the fragmentation pattern (MS/MS) for each peptide.
To get the highest sequence coverage possible, it is important to select the right proteases. Therefore, the first step is to perform bioinformatics analysis of your specific protein to advise on the type and number of proteases to apply for your project. This way, full sequence coverage is achieved, and you receive a detailed protein sequencing report.
Experience shows that many customers also combine peptide mapping with other services. Therefore, you may want to add N-terminal sequencing and intact mass analysis to extend the protein characterization.