Detection of Disulfide-Linked Peptides by HPLC

  • Alastair Aitken
  • Michèle Learmonth
Part of the Springer Protocols Handbooks book series (SPH)


Classical techniques for determining disulfide bond patterns usually require the fragmentation of proteins into peptides under low-pH conditions to prevent disulphide exchange. Pepsin or cyanogen bromide are particularly useful (see Chapter 63). Diagonal techniques to identify disulphide-linked peptides were developed by Brown and Hartley (see Chapter 81). A modern micromethod employing reverse-phase HPLC is described here.


Cyanogen Bromide Iodoacetic Acid Peptide Bond Cleavage Reverse Phase HPLC Column Reductive Alkylation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Friedman, M., Zahnley, J. C., and Wagner, J. R. (1980) Estimation of the disulfide content of trypsin inihibitors as S-β-(2-pyridylethyl)-L-cysteine. Anal. Biochem. 106, 27–34.PubMedCrossRefGoogle Scholar
  2. 2.
    Amons, R. (1987) Vapor-phase modification of sulfhydryl groups in proteins FEBS Lett. 212, 68–72.PubMedCrossRefGoogle Scholar
  3. 3.
    Toren, P., Smith, D., Chance, R., and Hoffman, J. (1988) Determination of interchain crosslinkages in insulin B-chain dimers by fast atom bombardment mass spectrometry. Anal. Biochem. 169, 287–299.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Alastair Aitken
    • 1
  • Michèle Learmonth
    • 1
  1. 1.National Institute of Medical ResearchMill Hill, LondonUK

Personalised recommendations