Recombinant proteinase 3 (Wregener's antigen) expressed in Pichia pastoris is functionally active and is recognized by patient seraHarmsen, MC., Heeringa, P., VanderGeld, YM., Huitema, MG., Klimp, A., Tiran, A. & Kallenberg, CGM., Nov-1997, In : Clinical and Experimental Immunology. 110, 2, p. 257-264 8 p.
Research output: Contribution to journal › Article › Academic › peer-review
The open reading frame of human proteinase 3 (PR3) without the prepro-peptide was cloned and expressed in Escherichia coli (rcPR3) and in Pichia pastoris (rpPR3). The 6-histidine tagged rpPR3 was efficiently secreted into culture supernatant from which it could be purified by immobilized metal chelate chromatography. Purified rpPR3 migrated as a single 32-kD band on SDS-PAGE and harboured protease activity that could be inhibited with inhibitors specific br serine-proteases. By indirect antigen-capture ELISA using rpPR3, 60% of sera from patients with Wegener's granulomatosis bound to the recombinant product, although it was not recognized in ELISA with directly coated rpPR3.
|Number of pages||8|
|Journal||Clinical and Experimental Immunology|
|Publication status||Published - Nov-1997|
- Wegner's granulomatosis, recombinant proteinase 3, human anti-neutrophil cytoplasmic antibody, Pichia, expression system, NEUTROPHIL SERINE PROTEINASE, ANTINEUTROPHIL CYTOPLASMIC AUTOANTIBODIES, WEGENER GRANULOMATOSIS AUTOANTIBODIES, DISEASE-ACTIVITY, ANTICYTOPLASMIC ANTIBODIES, LEUKOCYTE PROTEINASE, METHYLOTROPHIC YEAST, AUTOANTIGEN, PR3, INHIBITION