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High Level Production of Hybrid Poty virus–like Particles Carrying Repetitive Copies of Foreign Antigens in Escherichia coli

Bio/Technology volume 11, pages 1166–1170 (1993)Cite this article

Abstract

Synthesis in E. coli of native coat protein of Johnsongrass mosaic virus, and hybrid protein molecules containing foreign antigens, resulted in the intracellular formation of potyvirus-like particles (PVLPs). The foreign antigens used were an octapeptide epitope from Plasmodium falciparum and a decapeptide hormone (luetinizing hormone releasing hormone) at the N- or at both N- and C- terminal regions of the coat protein molecule, and a full length protein antigen (Sj26-glutathione S-transferase of 26 kD from Schistosoma japonicum) replacing the N- terminal 62 amino acids of the coat protein. Electron microscopy of ultrathin sections of E. coli revealed that PVLPs resulting from coat protein molecules containing peptide fusions appeared in vast arrays of parallel strands within the cytoplasm sometimes extending the length of the cell and at times the cells were strung together, with “threads” of PVLPs appearing to connect individual bacterial cells. PVLPs resulting from the fusion of the 26 kD antigen Sj26 to coat protein were shorter and wider. The physical form of the high molecular weight PVLPs enabled purification by simple size exclusion column chromatography. The Sj26-PVLPs administered to mice without adjuvant elicited antibody responses comparable to monomeric Sj26 administered with Freund's Complete Adjuvant.

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Author notes

  1. Mittur N. Jagadish: Corresponding author.

Authors and Affiliations

  1. CSIRO, Division of Biomolecular Engineering, 343 Royal Parade, Parkville, 3052, Victoria, Australia

    Mittur N. Jagadish, Caroline S. Fernandez & Colin W. Ward

  2. CSL Ltd., 45 Poplar Road, Parkville, 3052, Victoria, Australia

    Ross C. Hamilton, Peter Schoofs, Harry Kalnins & Ian T. Nisbet

  3. The Walter and Eliza Hall Institute of Medical Research, Parkville, 3052, Victoria, Australia

    Kathy M. Davern

Authors
  1. Mittur N. Jagadish
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  2. Ross C. Hamilton
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  4. Peter Schoofs
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Jagadish, M., Hamilton, R., Fernandez, C. et al. High Level Production of Hybrid Poty virus–like Particles Carrying Repetitive Copies of Foreign Antigens in Escherichia coli. Nat Biotechnol 11, 1166–1170 (1993). https://doi.org/10.1038/nbt1093-1166

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