Expression of the human autoantigen of Wegener's granulomatosis (PR3) in a baculovirus expression system

C. H. Szymkowiak; T. W. Johnston; E. Csernok; W. L. Gross

doi:10.1006/bbrc.1996.0224

Expression of the human autoantigen of Wegener's granulomatosis (PR3) in a baculovirus expression system

C. H. Szymkowiak, T. W. Johnston, E. Csernok, W. L. Gross^*

^*Corresponding author for this work

Department of Rheumatology and Clinical Immunology

21 Citations (Scopus)

Abstract

Proteinase 3 (PR3) is a multifunctional serine protease found in myeloperoxidase-positive granula of PMN. It is also the main target antigen for antineutrophil cytoplasmic antibodies (ANCA) found in Wegener's granulomatosis (WG). There is ample evidence that these autoantibodies play an active role in the pathogenesis of this disease. We have established the expression of the autoantigen (PR3) in a baculovirus expression system using insect cells. Recombinant PR3 was identified by IIF, direct ELISA using 3 different MoAbs and by catching ELISA using a MoAb and a rat polyclonal Ab or cANCA-positive WG patient sera. As purification of human PR3 from blood-derived PMNs has been shown to be difficult and very inefficient, insect cell-derived recombinant PR3 offers new possibilities for obtaining sufficient amounts of this protein to facilitate further biochemical characterizations.

Original language	English
Journal	Biochemical and Biophysical Research Communications
Volume	219
Issue number	2
Pages (from-to)	283-289
Number of pages	7
ISSN	0006-291X
DOIs	https://doi.org/10.1006/bbrc.1996.0224
Publication status	Published - 15.02.1996

Funding

The authors thank Mrs. Mons for her expert technical assistance. This work was supported by grants from the Bundes-ministerium für Forschung und Technologie (grant no.: 01 KD 89030), Bonn and from The Royal Society (grant no.: 418 GBR 112/5/94), London. Funds were also provided by the Verein zur Förderung der Erforschung und Bekämpfung rheumatischer Erkrankungen e. V., Bad Bramstedt, Germany.

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This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

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10.1006/bbrc.1996.0224

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title = "Expression of the human autoantigen of Wegener's granulomatosis (PR3) in a baculovirus expression system",

abstract = "Proteinase 3 (PR3) is a multifunctional serine protease found in myeloperoxidase-positive granula of PMN. It is also the main target antigen for antineutrophil cytoplasmic antibodies (ANCA) found in Wegener's granulomatosis (WG). There is ample evidence that these autoantibodies play an active role in the pathogenesis of this disease. We have established the expression of the autoantigen (PR3) in a baculovirus expression system using insect cells. Recombinant PR3 was identified by IIF, direct ELISA using 3 different MoAbs and by catching ELISA using a MoAb and a rat polyclonal Ab or cANCA-positive WG patient sera. As purification of human PR3 from blood-derived PMNs has been shown to be difficult and very inefficient, insect cell-derived recombinant PR3 offers new possibilities for obtaining sufficient amounts of this protein to facilitate further biochemical characterizations.",

author = "Szymkowiak, \{C. H.\} and Johnston, \{T. W.\} and E. Csernok and Gross, \{W. L.\}",

note = "Funding Information: The authors thank Mrs. Mons for her expert technical assistance. This work was supported by grants from the Bundes-ministerium f{\"u}r Forschung und Technologie (grant no.: 01 KD 89030), Bonn and from The Royal Society (grant no.: 418 GBR 112/5/94), London. Funds were also provided by the Verein zur F{\"o}rderung der Erforschung und Bek{\"a}mpfung rheumatischer Erkrankungen e. V., Bad Bramstedt, Germany. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.",

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T1 - Expression of the human autoantigen of Wegener's granulomatosis (PR3) in a baculovirus expression system

AU - Szymkowiak, C. H.

AU - Johnston, T. W.

AU - Csernok, E.

AU - Gross, W. L.

N1 - Funding Information: The authors thank Mrs. Mons for her expert technical assistance. This work was supported by grants from the Bundes-ministerium für Forschung und Technologie (grant no.: 01 KD 89030), Bonn and from The Royal Society (grant no.: 418 GBR 112/5/94), London. Funds were also provided by the Verein zur Förderung der Erforschung und Bekämpfung rheumatischer Erkrankungen e. V., Bad Bramstedt, Germany. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.

PY - 1996/2/15

Y1 - 1996/2/15

N2 - Proteinase 3 (PR3) is a multifunctional serine protease found in myeloperoxidase-positive granula of PMN. It is also the main target antigen for antineutrophil cytoplasmic antibodies (ANCA) found in Wegener's granulomatosis (WG). There is ample evidence that these autoantibodies play an active role in the pathogenesis of this disease. We have established the expression of the autoantigen (PR3) in a baculovirus expression system using insect cells. Recombinant PR3 was identified by IIF, direct ELISA using 3 different MoAbs and by catching ELISA using a MoAb and a rat polyclonal Ab or cANCA-positive WG patient sera. As purification of human PR3 from blood-derived PMNs has been shown to be difficult and very inefficient, insect cell-derived recombinant PR3 offers new possibilities for obtaining sufficient amounts of this protein to facilitate further biochemical characterizations.

AB - Proteinase 3 (PR3) is a multifunctional serine protease found in myeloperoxidase-positive granula of PMN. It is also the main target antigen for antineutrophil cytoplasmic antibodies (ANCA) found in Wegener's granulomatosis (WG). There is ample evidence that these autoantibodies play an active role in the pathogenesis of this disease. We have established the expression of the autoantigen (PR3) in a baculovirus expression system using insect cells. Recombinant PR3 was identified by IIF, direct ELISA using 3 different MoAbs and by catching ELISA using a MoAb and a rat polyclonal Ab or cANCA-positive WG patient sera. As purification of human PR3 from blood-derived PMNs has been shown to be difficult and very inefficient, insect cell-derived recombinant PR3 offers new possibilities for obtaining sufficient amounts of this protein to facilitate further biochemical characterizations.

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SN - 0006-291X

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JO - Biochemical and Biophysical Research Communications

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Expression of the human autoantigen of Wegener's granulomatosis (PR3) in a baculovirus expression system

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