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Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow
10:33

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Published on: January 8, 2014

A preliminary study for isolation of catalytic antibodies by histidine ligand affinity chromatography as an

E Nedonchelle1, O Pitiot, M A Vijayalakshmi

  • 1Laboratoire d'Interactions Moleculaires et de Technologie des Séparations (LIMTechS), Unité CNRS UPRES A 6022, Centre de Recherche de Royallieu, Compiègne, France.

Applied Biochemistry and Biotechnology
|May 29, 2000
PubMed

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-methylcoumarinamide 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.

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