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Autoantibodies pp 239-242 | Cite as

Assessment of Peptidylarginine Deiminase Activity by ELISA Using Human Fibrinogen as Substrate

  • Dres Damgaard
  • Claus H. Nielsen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1901)

Abstract

An enzyme-linked immunosorbent assay (ELISA) for measurement of the activity of peptidylarginine deiminases (PADs), the enzymes responsible for citrullination, is described. It uses fibrinogen as substrate for the enzyme and a commercial antibody specific for the citrullinated form of fibrinogen.

Key words

Citrullination ELISA Fibrinogen Peptidylarginine deiminase Rheumatoid arthritis 

References

  1. 1.
    Damgaard D, Senolt L, Nielsen M, Pruijn G, Nielsen CH (2014) Demonstration of extracellular peptidylarginine deiminase (PAD) activity in synovial fluid of patients with rheumatoid arthritis using a novel assay for citrullination of fibrinogen. Arthritis Res Ther 16(6):498CrossRefGoogle Scholar
  2. 2.
    Fujisaki M, Sugawara K (1981) Properties of peptidylarginine deiminase from the epidermis of newborn rats. J Biochem 89:257–263CrossRefGoogle Scholar
  3. 3.
    Nakayama-Hamada M, Suzuki A, Kubota K, Takazawa T, Ohsaka M, Kawaida R et al (2005) Comparison of enzymatic properties between hPADI2 and hPADI4. Biochem Biophys Res Commun 327:192–200CrossRefGoogle Scholar
  4. 4.
    Damgaard D, Senolt L, Nielsen CH (2016) Increased levels of peptidylarginine deiminase 2 in synovial fluid from anti-CCP-positive rheumatoid arthritis patients: association with disease activity and inflammatory markers. Rheumatology (Oxford) 55:918–927CrossRefGoogle Scholar
  5. 5.
    Damgaard D, Bjorn ME, Jensen PO, Nielsen CH (2017) Reactive oxygen species inhibit catalytic activity of peptidylarginine deiminase. J Enzyme Inhib Med Chem 32:1203–1208CrossRefGoogle Scholar
  6. 6.
    Damgaard D, Bjorn ME, Steffensen MA, Pruijn GJ, Nielsen CH (2016) Reduced glutathione as a physiological co-activator in the activation of peptidylarginine deiminase. Arthritis Res Ther 18:102CrossRefGoogle Scholar
  7. 7.
    Sipila KH, Ranga V, Rappu P, Torittu A, Pirila L, Kapyla J et al (2016) Extracellular citrullination inhibits the function of matrix associated TGF-beta. Matrix Biol 55:77–89CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Dres Damgaard
    • 1
    • 2
  • Claus H. Nielsen
    • 1
    • 2
  1. 1.Institute for Inflammation Research, Center for Rheumatology and Spine DiseasesCopenhagen University Hospital, RigshospitaletCopenhagenDenmark
  2. 2.Section for Periodontology, Microbiology and Community Dentistry, Department of Odontology, Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagenDenmark

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