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Expression of Active, Secreted Human Prostate-specific Antigen by Recombinant Baculovirus-infected Insect Cells on a Pilot-scale

Bio/Technology volume 13, pages 1230–1234 (1995)Cite this article

Abstract

We have expressed human prostate-specific antigen (PSA) on a pilot-scale in Spodoptera frugiperda Sf9 insect cells using recombinant baculovirus system. Infected cells secreted PSA into culture medium at a concentration of 2–4 mg per liter. PSA was expressed both in active and inactive forms which were separated in a final purification step using cation-exchange chromatography eluted with a low salt gradient. The N-terminus of active PSA was correctly cleaved; two amino acids of the propeptide remained, however, at the N-terminus of the inactive PSA. Purified recombinant PSA showed a chymotrypsin-like activity with the synthetic substrate MeO-Suc-Arg-Pro-Tyr-pNA, but did not have a trypsin-like activity when Pro-Phe-Arg-pNA was used. The molecular mass of active PSA was 31.0 kDa in reduced SDS-PAGE, 26.0 kDa in nonreduced SDS-PAGE and 26.5 kDa in ion spray mass spectrometry. The active protein formed complexes with α1-antichymotrypsin (ACT) and α2-macroglobulin (α2M) in vitro similar to the commercial PSA purified from human seminal fluid.

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  1. Pirkko Vihko: Corresponding author.

Authors and Affiliations

  1. Biocenter Oulu and Department of Clinical Chemistry, University of Oulu, FIN-90220, Oulu, Finland

    Riitta Kurkela, Annakaisa Herrala, Pirkko Henttu, Heini Nal & Pirkko Vihko

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  1. Riitta Kurkela
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Kurkela, R., Herrala, A., Henttu, P. et al. Expression of Active, Secreted Human Prostate-specific Antigen by Recombinant Baculovirus-infected Insect Cells on a Pilot-scale. Nat Biotechnol 13, 1230–1234 (1995). https://doi.org/10.1038/nbt1195-1230

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