| Title | A preliminary study for isolation of catalytic antibodies by histidine ligand affinity chromatography as an alternative to conventional protein A/G methods. | | Author(s) | Nedonchelle E, Pitiot O, Vijayalakshmi MA | | Institution | Laboratoire d'Interactions Moleculaires et de Technologie des Séparations (LIMTechS), Unité CNRS UPRES A 6022, Centre de Recherche de Royallieu, Compiègne, France. | | Source | Appl Biochem Biotechnol 2000 Jan-Mar; 83(1-3):287-94; discussion 294-5, 297-313. | | MeSH | Antibodies, Catalytic
Autoantibodies
Chromatography, Affinity
Comparative Study
Histidine
Humans
Hydrolysis
Immunoglobulin G
In Vitro
Ligands
Lupus Erythematosus, Systemic
Nerve Tissue Proteins
Oligopeptides
Research Support, Non-U.S. Gov't
Staphylococcal Protein A
| | 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-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. | | Language | eng | | Pub Type(s) | Journal Article
| | PubMed ID | 10826967 |
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