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Adsorption Study of Immunoglobulin G Subclasses from Different Species by Pseudobioaffinity Separation on Histidyl–Bisoxirane–Sepharose

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Abstract

Adsorption chromatography is increasingly used for protein separations and biomedical applications. Therapeutic molecules such as antibodies, cytokines, therapeutic DNA, and plasma proteins must be purified before characterization and utilization. Use of immunoglobulins as immunodiagnostic and therapeutic tools has initiated many attempts to develop new adsorbents for their separation. Protein A and protein G are the affinity ligands most widely used for separation of immunoglobulins. These proteins are reliable, and have good selectivity and specificity, but are very expensive. Much attention has therefore been devoted to developing alternative methods for separation of immunoglobulins. Pseudobiospecific ligands, for example metal ions and amino acids, can be used for separation of a wide range of biological molecules. In this study, IgG1, IgG2, and IgG3, three subclasses of human IgG, were separated from human serum using the amino acid histidine grafted on to bisoxirane-activated Sepharose, as pseudobiospecific adsorbent. Adsorption of IgG from different animal species on the same chromatographic adsorbent was also tested. The high recovery and purification on histidyl–bisoxirane–Sepharose gel of IgG from all the sources tested compared well with results obtained by use of protein A–Sepharose gel.

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Acknowledgments

This work was supported by a research grant provided by the Lebanese National Council for Scientific Research (LNCSR; project No MD. 579).

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Authors and Affiliations

  1. Laboratoire de Bioséparation et d’Interactions Moléculaires (BiosIM), Département de Biologie Médicale, Faculté de Pharmacie, Université Libanaise, 14/6573, Beirut, Lebanon

    Assem Elkak & Sanaa Ismail

  2. The National Council for Scientific Research, 11/8281, Beirut, Lebanon

    Assem Elkak

  3. Al-Zahraa University Hospital, Aljinah, Beirut, Lebanon

    Sanaa Ismail

  4. Department of Chemistry, Biochemistry Division, Hacettepe University, Beytepe, Ankara, 06242, Turkey

    Lokman Uzun & Adil Denizli

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  1. Assem Elkak
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  2. Sanaa Ismail
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Correspondence to Assem Elkak.

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Elkak, A., Ismail, S., Uzun, L. et al. Adsorption Study of Immunoglobulin G Subclasses from Different Species by Pseudobioaffinity Separation on Histidyl–Bisoxirane–Sepharose. Chroma 69, 1161–1167 (2009). https://doi.org/10.1365/s10337-009-1071-6

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  • DOI: https://doi.org/10.1365/s10337-009-1071-6

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