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Rapid metal-interaction chromatography of proteins and peptides on micropellicular sorbents

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Summary

Short columns packed with micropellicular stationary phases consisting of 2-μm fused silica microspheres with covalently bound iminodiacetate (IDA) functions at the surface were used for rapid HPLC analysis of proteins by metal-interaction chromatography (MIC). In contrast to conventional porous stationary phases which elicit relatively long analysis times, the columns packed with sorbents having micropellicular configuration and Ni2+ or Co2+ chelated by the IDA functions yielded separation of model proteins in a few minutes with good resolution. A Fe3+/IDA column was used for separation of phosphorylated and non-phosphorylated peptides derived from enzymatically digested erythrocyte membrane proteins. Stability of the Fe3+/IDA column was quite satisfactory as determined by monitoring the iron content of the column effluent and by measuring the amount of iron present in the stationary phase.

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Authors and Affiliations

  1. Institute of Radiochemistry, University of Innsbruck, A-6020, Innsbruck, Austria

    G. K. Bonn

  2. Department of Chemical Engineering, Yale University, 06520, New Haven, CT, USA

    K. Kalghatgi & Cs. Horváth

  3. Department of Pathology, School of Medicine, Yale University, 06520, New Haven, CT, USA

    W. C. Horne

Authors
  1. G. K. Bonn
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  2. K. Kalghatgi
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  3. W. C. Horne
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  4. Cs. Horváth
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Bonn, G.K., Kalghatgi, K., Horne, W.C. et al. Rapid metal-interaction chromatography of proteins and peptides on micropellicular sorbents. Chromatographia 30, 484–488 (1990). https://doi.org/10.1007/BF02269792

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  • DOI: https://doi.org/10.1007/BF02269792

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