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Catalytic and DNA-Hydrolyzing Activities of Purified Immunoglobulins IgG from Patients Sera with Autoimmune Disease by Pseudobiospecific Chromatography Using Histidyl-Aminohexyl-Sepharose
Authors:A Elkak  M Bourhim  Y Coffinier  M A Vijayalakshmi
Institution:(1) Department of Medical Biology, Faculty of Pharmacy, Lebanese University, P.O.Box. 14/6573, Beirut, Lebanon;(2) The National Council for Scientific Research, P.O.Box. 11/8281, Beirut, Lebanon;(3) Laboratoire drsquoInteractions Moléculaires et de Technologie des séparations, Université de Technologie de Compiègne, B.P. 20569, 60205 Compiègne, France
Abstract:Catalytic autoimmune antibodies from patients with antiphospholipid (aPL) antibodies were purified using histidyl-aminohexyl-sepharose gel. The sera were loaded on the columns equilibrated with 25 mM MOPS buffer pH 7.4 and the absorbed proteins were eluted by adding 0.2 M NaCl in the equilibrating buffer. Antibodies purity was evaluated by SDS-PAGE. The purified immunoglobulins G from patients with (aPL) sera by histidyl-aminohexyl-sepharose show DNA-degrading activity of the plasmid pUC19 DNA and catalytic activity in hydrolyzing the peptide substrate Pro-Phe-Arg-7-amido-4-methylcoumarin.
Keywords:Column liquid chromatography  Pseudobioaffinity chromatography  Histidine-aminohexyl-sepharose gel  Autoimmune diseases  DNA-hydrolysis
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