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Recombinant Protein Expression in Nicotiana

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Plant Chromosome Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 701))

Abstract

Recombinant protein pharmaceuticals are now widely used in treatment of chronic diseases, and several recombinant protein subunit vaccines are approved for human and veterinary use. With growing demand for complex protein pharmaceuticals, such as monoclonal antibodies, manufacturing capacity is becoming limited. There is increasing need for safe, scalable, and economical alternatives to mammalian cell culture-based manufacturing systems, which require substantial capital investment for new manufacturing facilities. Since a seminal paper reporting immunoglobulin expression in transgenic plants was published in 1989, there have been many technological advances in plant expression systems to the present time where production of proteins in leaf tissues of nonfood crops such as Nicotiana species is considered a viable alternative. In particular, transient expression systems derived from recombinant plant viral vectors offer opportunities for rapid expression screening, construct optimization, and expression scale-up. Extraction of recombinant proteins from Nicotiana leaf tissues can be achieved by collection of secreted protein fractions, or from a total protein extract after grinding the leaves with buffer. After separation from solids, the major purification challenge is contamination with elements of the photosynthetic complex, which can be solved by application of a variety of facile and proven strategies. In conclusion, the technologies required for safe, efficient, scalable manufacture of recombinant proteins in Nicotiana leaf tissues have matured to the point where several products have already been tested in phase I clinical trials and will soon be followed by a rich pipeline of recombinant vaccines, microbicides, and therapeutic proteins.

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Acknowledgement

We thank Andrew Marsh for editorial assistance.

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Authors and Affiliations

  1. Owensboro Cancer Research Program, Owensboro, KY, USA

    Nobuyuki Matoba, Keith R. Davis & Kenneth E. Palmer

  2. Department of Pharmacology & Toxicology and the James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY, USA

    Nobuyuki Matoba, Keith R. Davis & Kenneth E. Palmer

Authors
  1. Nobuyuki Matoba
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  2. Keith R. Davis
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  3. Kenneth E. Palmer
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Editor information

Editors and Affiliations

  1. , Division of Biological Sciences, University of Missouri, Tucker Hall 117, Columbia, 65211, Missouri, USA

    James A. Birchler

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Matoba, N., Davis, K.R., Palmer, K.E. (2011). Recombinant Protein Expression in Nicotiana . In: Birchler, J. (eds) Plant Chromosome Engineering. Methods in Molecular Biology, vol 701. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-957-4_11

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  • DOI: https://doi.org/10.1007/978-1-61737-957-4_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61737-956-7

  • Online ISBN: 978-1-61737-957-4

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