Isolation of high-specific-activity subunits of cholera toxin by reversed-phase high-performance liquid chromatography

Stephen D. Pearson, James D. Dixon, Steven F. Northwehr, Alexander Kurosky

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    4 Scopus citations

    Abstract

    Facile, rapid procedures for the separation of active cholera toxin subunits were developed, based on high-performance liquid chromatography (HPLC) with a Nucleosil C8 reversed-phase column. These procedures were capable of completely resolving subunits A and B as well as S-carboxymethylated or reduced α-, γ-, and β-chains. The binding of HPLC-purified B subunit to GM1 ganglioside was essentially identical to that of cholera toxin when compared on a molar basis. The adenosine 5′-diphosphate-ribosyltransferase activity of HPLC-purified A subunit, reduced α-chain, or carboxymethyl α-chain was also determined to be reasonably high compared to that of cholera toxin or commercially prepared A subunit.

    Original languageEnglish (US)
    Pages (from-to)413-421
    Number of pages9
    JournalJournal of Chromatography A
    Volume359
    Issue numberC
    DOIs
    StatePublished - 1986

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    ASJC Scopus subject areas

    • Analytical Chemistry
    • Clinical Biochemistry
    • Molecular Medicine

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