Glyco-analytical multispecific proteolysis (Glyco-AMP): A simple method for detailed and quantitative glycoproteomic characterization

  • Hua S
  • Hu C
  • Kim B
 et al. 
  • 13

    Readers

    Mendeley users who have this article in their library.
  • 25

    Citations

    Citations of this article.

Abstract

Despite recent advances, site-specific profiling of protein glycosylation remains a significant analytical challenge for conventional proteomic methodology. To alleviate the issue, we propose glyco-analytical multispecific proteolysis (Glyco-AMP) as a strategy for glycoproteomic characterization. Glyco-AMP consists of rapid, in-solution digestion of an analyte glycoprotein (or glycoprotein mixture) by a multispecific protease (or protease cocktail). Resulting glycopeptides are chromatographically separated by isomer-specific porous graphitized carbon nano-LC, quantified by high-resolution MS, and structurally elucidated by MS/MS. To demonstrate the consistency and customizability of Glyco-AMP methodology, the glyco-analytical performances of multispecific proteases subtilisin, pronase, and proteinase K were characterized in terms of quantitative accuracy, sensitivity, and digestion kinetics. Glyco-AMP was shown be effective on glycoprotein mixtures as well as glycoproteins with multiple glycosylation sites, providing detailed, quantitative, site- and structure-specific information about protein glycosylation.

Author-supplied keywords

  • biopharmaceutical glycoproteins
  • glycan isomers
  • glycoproteomics
  • multispecific proteases
  • nonspecific proteases
  • site-specific glycosylation

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Get full text

Authors

  • Serenus Hua

  • Chloe Y. Hu

  • Bum Jin Kim

  • Sarah M. Totten

  • Myung Jin Oh

  • Nayoung Yun

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free