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Applied Biochemistry and Biotechnology

, Volume 83, Issue 1–3, pp 287–295 | Cite as

A preliminary study for isolation of catalytic antibodies by histidine ligand affinity chromatography as an alternative to conventional protein A/G methods

  • Elsa Nedonchelle
  • Olivier Pitiot
  • Mookambeswaran A. Vijayalakshmi
Article

Abstract

Catalytic autoimmune antibodies from the sera of lupus patients were purified using histidyl-aminohexyl-Sepharose gel and compared with the antibodies purified with protein A and protein G affinity chromatography. The IgG preparations from the histidine affinity column had a much higher catalytic activity in hydrolyzing the peptide substrate Pro-Phe-Arg-methyl-coumarinamide compared to the antibodies obtained by the conventional protein A/G method. This preservation of catalytic activity is attributed to the gentle buffer conditions used in the histidine ligand method that allowed the integrity of three-dimensional structure of purified catalytic antibodies. Thus, histidine affinity offer a superior method for isolating autoimmune catalytic antibodies.

Index Entries

Catalytic autoimmune antibodies activity preservation immobilized histidine affinity chromatography 

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Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Elsa Nedonchelle
    • 1
  • Olivier Pitiot
    • 1
  • Mookambeswaran A. Vijayalakshmi
    • 1
  1. 1.Laboratoire d'Interactions Moleculaires et de Technologie des Séparations (LIMTechS)Unité CNRS UPRES A 6022, Centre de Recherche de RoyallieuCompiègne cedexFrance

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