Jump to main content
Jump to site search

Issue 5, 2017
Previous Article Next Article

Detection of glycosylation and iron-binding protein modifications using Raman spectroscopy

Author affiliations

Abstract

In this study we demonstrate the use of Raman spectroscopy to determine protein modifications as a result of glycosylation and iron binding. Most proteins undergo some modifications after translation which can directly affect protein function. Identifying these modifications is particularly important in the production of biotherapeutic agents as they can affect stability, immunogenicity and pharmacokinetics. However, post-translational modifications can often be difficult to detect with regard to the subtle structural changes they induce in proteins. From their Raman spectra apo-and holo-forms of iron-binding proteins, transferrin and ferritin, could be readily distinguished and variations in spectral features as a result of structural changes could also be determined. In particular, differences in solvent exposure of aromatic amino acids residues could be identified between the open and closed forms of the iron-binding proteins. Protein modifications as a result of glycosylation can be even more difficult to identify. Through the application of the chemometric techniques of principal component analysis and partial least squares regression variations in Raman spectral features as a result of glycosylation induced structural modifications could be identified. These were then used to distinguish between glycosylated and non-glycosylated transferrin and to measure the relative concentrations of the glycoprotein within a mixture of the native non-glycosylated protein.

Graphical abstract: Detection of glycosylation and iron-binding protein modifications using Raman spectroscopy

Back to tab navigation

Publication details

The article was received on 22 Nov 2016, accepted on 01 Feb 2017 and first published on 01 Feb 2017


Article type: Paper
DOI: 10.1039/C6AN02516A
Citation: Analyst, 2017,142, 808-814
  • Open access: Creative Commons BY license
  •   Request permissions

    Detection of glycosylation and iron-binding protein modifications using Raman spectroscopy

    L. Ashton, V. L. Brewster, E. Correa and R. Goodacre, Analyst, 2017, 142, 808
    DOI: 10.1039/C6AN02516A

    This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.

    Reproduced material should be attributed as follows:

    • For reproduction of material from NJC:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the Centre National de la Recherche Scientifique (CNRS) and the RSC.
    • For reproduction of material from PCCP:
      [Original citation] - Published by the PCCP Owner Societies.
    • For reproduction of material from PPS:
      [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
    • For reproduction of material from all other RSC journals:
      [Original citation] - Published by The Royal Society of Chemistry.

    Information about reproducing material from RSC articles with different licences is available on our Permission Requests page.

Search articles by author

Spotlight

Advertisements