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

    49 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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