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Binding site on human immunoglobulin G for the affinity ligand HWRGWV

Authors

  • Haiou Yang,

    1. Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA
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  • Patrick V. Gurgel,

    1. Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA
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  • D. Keith Williams Jr,

    1. W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA
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  • Benjamin G. Bobay,

    1. Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA
    2. North Carolina Research Campus, Kannapolis, Kannapolis, NC 28081, USA
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  • John Cavanagh,

    1. Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA
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  • David C. Muddiman,

    1. W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA
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  • Ruben G. Carbonell

    Corresponding author
    1. Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA
    • Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA.
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Abstract

Affinity ligand HWRGWV has demonstrated the ability to isolate human immunoglobulin G (hIgG) from mammalian cell culture media. The ligand specifically binds hIgG through its Fc portion. This work shows that deglycosylation of hIgG has no influence on its binding to the HWRGWV ligand and the ligand does not compete with Protein A or Protein G in binding hIgG. It is suggested by the mass spectrometry (MS) data and docking simulation that HWRGWV binds to the pFc portion of hIgG and interacts with the amino acids in the loop Ser383–Asn389 (SNGQPEN) located in the CH3 domain. Subsequent modeling has suggested a possible three-dimensional minimized solution structure for the interaction of hIgG and the HWRGWV ligand. The results support the fact that a peptide as small as a hexamer can have specific interactions with large proteins such as hIgG. Copyright © 2009 John Wiley & Sons, Ltd.

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