Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis

AA Rarok, MG Huitema, MJ van der Leij, YM van der Geld, H Berthold, J Schmitt, CA Stegeman, PC Limburg, CGM Kallenberg*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen.

Original languageEnglish
Pages (from-to)586-595
Number of pages10
JournalAMERICAN JOURNAL OF CLINICAL PATHOLOGY
Volume120
Issue number4
DOIs
Publication statusPublished - Oct-2003

Keywords

  • recombinant PR3
  • proteinase 3
  • PR3-ANCA
  • antineutrophil cytoplasmic antibodies
  • systemic vasculitis
  • ELISA
  • enzyme-linked immunosorbent assay
  • ANTINEUTROPHIL CYTOPLASMIC ANTIBODIES
  • WEGENERS-GRANULOMATOSIS
  • CRESCENTIC GLOMERULONEPHRITIS
  • MONOCLONAL-ANTIBODIES
  • DISEASE-ACTIVITY
  • CAPTURE-ELISA
  • C-ANCA
  • PR3
  • AUTOANTIGEN
  • EXPRESSION

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