C.A.T. GmbH & Co. Chromatographie und Analysentechnik KG

Heerweg 10
D-72070 Tübingen
Germany

Phone: +49 (0) 7071 / 78323
Fax: +49 (0) 7071 / 75163

General E-Mail:
info@cat-online.com

Personal E-Mails:
Team

Services > Sequencing

 

Sequencing

  • Sequencing
  • Peptide sequence via MSn
  • Cyclic peptides

For peptide sequencing we use the MS/MS method. Whether complete elucidation of the sequence is possible or not depends on the nature of the sequence. For peptides containing 20-30 amino acid residues, complete sequence elucidation is usually possible.

Difference between Edman and MSn sequencing:

Whereas Edman degradation starts at the N-terminus and proceeds sequentially to the C-terminus, MSn fragments the complete molecule. As a consequence:

Under certain circumstances Edman can fail after a couple of amino acids whereas

MS/MS can fail for longer peptides primarily at the N- and/or C-terminus.

In contrast to Edman degradation, MS/MS can fail only for a short part of the sequence but verifying the sequence on both sides of the unverified part.

SequenceDiagramm

Sequence

MSn or the combination HPLC-MSn can be offered for peptide sequencing, for the determination of the molecular mass of peptides and for the identification of side products. If necessary, separation can be performed using HPLC on suitable columns. Peaks are detected by diode array detector and transferred online into the mass spectrometer. After electro-spray ionization (ESI) the components are identified by MSn.

Results: Molecular mass y,
b and a-type fragments found.
The determined peptide sequence.

Sequenc3
sequenc4

Prior to sequencing of cyclic peptides with a disulfide bridge, the ring structure must be opened selectively.
After disulfide cleavage with DTT (1,4-Dithio-DL-threitol), the molecular weight is checked to be 2 mass units higher than for the cyclic peptide and the linear peptide is subsequently sequenced.

cyclic

Inspections

FDA Inspection

May 2015, successful (No 483)

German authorities

Juni 2015, successful


Symposium

Please visit our booth at:

34th European Peptide Symposium 2016, Leipzig

4th to 9th Septembre 2016

see Links