Collection of α1-acid glycoprotein molecular species by capillary electrophoresis and the analysis of their molecular masses and carbohydrate chains: Basic studies on the analysis of glycoprotein glycoforms

Kazuo Sei, Miyako Nakano, Mitsuhiro Kinoshita, Takashi Masuko, Kazuaki Kakehi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

48 Citations (Scopus)

Abstract

A highly heterogeneous glycoprotein, α1-acid glycoprotein, was resolved into their glycoforms by capillary electrophoresis using a surface-modified capillary in 20 mM acetate buffer (pH 4.2) containing 0.5% (w/v) hydroxypropylmethylcellulose. We collected the fractions containing each glycoform as nearly pure state by capillary electrophoresis, and examined the molecular masses of these glycoforms by matrix assisted laser desorption time-of-flight mass spectrometry. We also analyzed carbohydrate chains after releasing them with N-glycosidase F followed by fluorescent labeling with 8-aminopyrene-1,3,6-trisulfonate. We found that the separation of glycoforms was mostly due to the presence of multiantennary carbohydrate chains. We propose that the present technique is useful for the analysis of post translational modification of proteins with carbohydrate chains.

Original languageEnglish
Pages (from-to)273-281
Number of pages9
JournalJournal of Chromatography A
Volume958
Issue number1-2
DOIs
Publication statusPublished - 7 Jun 2002

Keywords

  • Alpha1-acid glycoprotein
  • Carbohydrates
  • Glycoproteins
  • Orosomucoid
  • Proteins

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