The determination of a protein's sequence is can be necessary especially when no DNA data is available. This is especially true for
- proteins from non-common species where no genome data exists
- monoclonal antibodies when no hybridoma cells are avalable
- commercial proteins with undisclosed sequence
Aim of de novo sequencing is either to obtain peptide sequence tags for the generation of DNA primers or the elucidation of the full protein sequence, e. g. for the generation of biosimilars.
To tackle such tasks, we initially apply
- different proteolytic digests of the protein
- high resolution mass spectrometry to generate high confidence data
- extensive bioinformatic analysis
If necessary further analyses can include
- micropreparative HPLC of proteolytic peptides
- Edman sequencing of internal fragments
- experimental validation of results via synthetic peptides
As the scope and complexity of each sequencing project will vary by prior knowledge of the protein, its size and primary structure, please contact our specialists for consultation.