Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis.

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.

journal_name

Am J Clin Pathol

authors

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

doi

10.1309/YTU2-FUHB-EXJL-WMLD

subject

Has Abstract

pub_date

2003-10-01 00:00:00

pages

586-95

issue

4

eissn

0002-9173

issn

1943-7722

journal_volume

120

pub_type

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