, Volume 60, Issue 3–4, pp 165–169 | Cite as

Catalytic and DNA-Hydrolyzing Activities of Purified Immunoglobulins IgG from Patients Sera with Autoimmune Disease by Pseudobiospecific Chromatography Using Histidyl-Aminohexyl-Sepharose

  • A. Elkak
  • M. Bourhim
  • Y. Coffinier
  • M. A. Vijayalakshmi


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.


Column liquid chromatography Pseudobioaffinity chromatography Histidine-aminohexyl-sepharose gel Autoimmune diseases DNA-hydrolysis 


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  1. Khamashta MA, Hughes GR (1994) Clin Rev Allergy 12:287–296PubMedGoogle Scholar
  2. Harris EN, Gharavi AE, Boey ML, Patel BM, Mackworth-Young C, Hughes GR (1983) Lancet 2:1211–1214CrossRefPubMedGoogle Scholar
  3. Greaves M (1999) Lancet 353:1348–1353CrossRefPubMedGoogle Scholar
  4. Fehr T, Cathomas G, Weber C,Fontana A, Schaffner A (2001) Lupus 10 (8):576–579Google Scholar
  5. Galli M, Finazzi G, Barbui T (1996) Br J Haemat 93 (1):1–5Google Scholar
  6. Tyutulkova S, Paul S (1994) Appl Biochem Biotech 47:191–198Google Scholar
  7. Tyutulkova S, Gao QS, Paul S (1995) In Methods in Molecular Biology. Antibody Engineering Protocols, Paul S (ed) Humana, Totowa NJ 51:377–394Google Scholar
  8. Buchner J, Renner M, Lilie H, Hin HJ, Jaenicke R (1991) Biochemistry 30:6922–6929PubMedGoogle Scholar
  9. Haupt K, Bueno SMA,Vijayalakshmi MA (1995) J Chromatogr B 674:13–21CrossRefGoogle Scholar
  10. Bueno SMA, Haupt K, Vijayalakshmi MA (1995) Int J Artif Organs 18 (70):392–398Google Scholar
  11. Pitiot O, Nedonchelle E, Legallais C, Vijayalakshmi MA (2001) J Chromatogr B 758 (2):173–182Google Scholar
  12. Shuster AM, Gololobov GV, Kvashuk OK, Bogomolova ATE, Smirnov IV, Gabibov AG (1992) Science 256:665–667PubMedGoogle Scholar
  13. Sarath G, De La Motte RS, Wagner FW (1989) In Proteolytic Enzymes: A Practical approach, Beyroun RJ, Bond JS (eds) Oxford UK: 25–55Google Scholar
  14. El-KAK A, Manjini S, Vijayalakshmi MA (1992) J Chromatogr 604:29–37CrossRefPubMedGoogle Scholar
  15. Nedonchelle E, Pitiot O, Vijayalakshmi MA (2000) Appl Biochem Biotech 83:287–294CrossRefGoogle Scholar
  16. El-KAK A, Vijayalakshmi MA (1992) Bioseparation 3:47–53PubMedGoogle Scholar

Copyright information

© Friedr. Vieweg&Sohn Verlagsgesellschaft mbH 2004

Authors and Affiliations

  • A. Elkak
    • 1
    • 2
  • M. Bourhim
    • 3
  • Y. Coffinier
    • 3
  • M. A. Vijayalakshmi
    • 3
  1. 1.Department of Medical BiologyFaculty of Pharmacy, Lebanese UniversityBeirutLebanon
  2. 2.The National Council for Scientific ResearchBeirutLebanon
  3. 3.Laboratoire d’Interactions Moléculaires et de Technologie des séparationsUniversité de Technologie de CompiègneCompiègneFrance

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