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Immobilized serum albumin: Rapid HPLC probe of stereoselective protein-binding interactions

Authors

  • Enrico Domenici,

    1. Centro di Studio del C.N.R. per le Macromolecole Stereordinate ed Otticamente Attive, Dipartimento di Chimica e Chimica Industriale, Universita di Pisa, Pisa, Italy
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  • Carlo Bertucci,

    1. Centro di Studio del C.N.R. per le Macromolecole Stereordinate ed Otticamente Attive, Dipartimento di Chimica e Chimica Industriale, Universita di Pisa, Pisa, Italy
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  • Piero Salvadori,

    1. Centro di Studio del C.N.R. per le Macromolecole Stereordinate ed Otticamente Attive, Dipartimento di Chimica e Chimica Industriale, Universita di Pisa, Pisa, Italy
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  • Sylvie Motellier,

    1. St. Jude Children's Research Hospital, Memphis, Tennessee 38105
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  • Irving W. Wainer

    Corresponding author
    1. St. Jude Children's Research Hospital, Memphis, Tennessee 38105
    • McGill University, Department of Oncology, 3655 Drummond, Suite 701, Montreal, PQ, Canada H36 1Y6
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Abstract

A human serum albumin-based HPLC chiral stationary phase (HSA-CSP) has been examined as a tool to investigate binding of chiral drugs to HSA and drug–drug protein-binding interactions. Rac-oxazepam hemisuccinate (OXH) was used as a model compound and the chromatographic retention (k′) of its enantiomers was determined after addition of displacers to the mobile phase. Compounds known to bind at the same site as OXH and at different sites were tested for their displacing capacities. Competitive binding interactions between the OXH enantiomers and displacers in the mobile phase were reflected by decreases in the k′s of (R)- and (S)-OXH. The results indicate that retention on the HSA-CSP accurately reflects binding to native HSA and the technique can determine enantioselective and competitive binding interactions at specific sites on HSA. The HSA-CSP was also able to recognize separate binding areas for (S)- and (R)-OXH.

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