A rapid ELISA based method to determine peptidyl-arginine deiminase activity in biological samples

  • S Nijenhuis1,

    Affiliated with

    • AJW Zendman1,

      Affiliated with

      • JMH Raats1,

        Affiliated with

        • GJM Pruijn1 and

          Affiliated with

          • WJ van Venrooij1

            Affiliated with

            Arthritis Res Ther20035(Suppl 1):17

            DOI: 10.1186/ar647

            Published: 24 February 2003

            Peptidylarginine deiminases (PADs; EC 3.5.3.15) are a family of calcium-dependent enzymes that convert peptidylarginine into peptidylcitrulline. The recent finding that patients with rheumatoid arthritis (RA) produce autoantibodies against citrulline-containing epitopes greatly increased the interest in the PAD enzymes and their activities. It is not yet known whether there is a causative relationship between the generation of antibodies targeting citrullinated epitopes and the development of the disease. Characterisation of the structure and function of PADs may help to understand the production process of citrullinated antigens and possibly also the aetiology of RA.

            Several assays are known to monitor PAD activity in biological samples. However, these assays either have a low sensitivity or are laborious. Here, we describe the development of a simple, rapid method for the simultaneous analysis of many PAD-containing samples. This new method is based on the binding of an antibody specifically recognising a citrulline-containing epitope in a defined peptide. We show that this method is very sensitive and can be applied to monitor PAD activity in many types of biological samples, such as bacterial lysates, mammalian cell extracts and tissue extracts.

            Authors’ Affiliations

            (1)
            Department of Biochemistry, University of Nijmegen

            Copyright

            © BioMed Central Ltd 2003

            Advertisement