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Expression and purification of an anti-Foot-and-mouth disease virus single chain variable antibody fragment in tobacco plants

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Abstract

Low-cost recombinant antibodies could provide a new strategy to control Foot-and-mouth disease virus (FMDV) outbreaks by passive immunization of susceptible animals. In this study, a single chain variable antibody fragment (scFv) recognizing FMDV coat protein VP1 was expressed in transgenic tobacco plants. To enhance the accumulation of scFv protein, the codon-usage of a murine hybridoma-derived scFv gene was adjusted to mimic highly expressed tobacco genes and fused to an elastin-like polypeptide (ELP) tag. This scFv–ELP fusion accumulated up to 0.8% of total soluble leaf protein in transgenic tobacco. To recover scFv–ELP protein from the leaf extract, a simple and scalable purification strategy was established. Purified scFv–ELP fusion was cleaved to separate the scFv portion. Finally, it was shown that the purified scFv proteins retained their capacity to bind the FMDV in the absence or presence of ELP fusion.

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Acknowledgments

The authors wish to thank Jamie McNeil, Lisa Starr, and the staff at the National Centre for Foreign Animal Disease for technical assistance. Alex Molnar is acknowledged for his assistance with the preparation of the figures. This research was supported by Agriculture and Agri-Food Canada’s Matching Investment Initiative program. The Academy of Finland is acknowledged for providing a fellowship for J.J.J., and the Natural Sciences and Engineering Research Council (NSERC) is thanked for providing financial support to A.J.C.

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  1. J. J. Joensuu

    Present address: VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Espoo, Finland

Authors and Affiliations

  1. Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON, N5V 4T3, Canada

    J. J. Joensuu, K. D. Brown, A. J. Conley, R. Menassa & J. E. Brandle

  2. Department of Biology, University of Western Ontario, London, ON, N6A 5B7, Canada

    A. J. Conley

  3. Canadian Food Inspection Agency, Winnipeg, MB, R3E 3M4, Canada

    A. Clavijo

  4. Vineland Research and Innovation Centre, Vineland Station, ON, L0R 2E0, Canada

    J. E. Brandle

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  1. J. J. Joensuu
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Correspondence to R. Menassa.

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Joensuu, J.J., Brown, K.D., Conley, A.J. et al. Expression and purification of an anti-Foot-and-mouth disease virus single chain variable antibody fragment in tobacco plants. Transgenic Res 18, 685–696 (2009). https://doi.org/10.1007/s11248-009-9257-0

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  • DOI: https://doi.org/10.1007/s11248-009-9257-0

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