Abstract
Transgenic plants for the production of high-value recombinant complex and/or glycosylated proteins are a promising alternative for conventional systems, such as mammalian cells and bacteria. Many groups use plants as production platform for antibodies and antibody fragments. Here, we describe how bivalent camel-like antibodies can be produced in leaves and seeds. Camel-like antibodies are fusions of the antigen-binding domain of heavy chain camel antibodies (VHH) with an Fc fragment of choice. Transient expression in Nicotiana benthamiana leaves allows the production of VHH-Fc antibodies within a few days after the expression plasmid has been obtained. Generation of stable Arabidopsis thaliana transformants allows production of scalable amounts of VHH-Fc antibodies in seeds within a year. Further, we describe how the in planta-produced VHH-Fc antibodies can be quantified by Western blot analysis with Fc-specific antibodies.
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Acknowledgments
The authors thank Martine De Cock for help in preparing the manuscript. This work is supported by the European Commission 6th Framework (Pharma Planta, LSHB-CT-2003-503565). We also thank the colleagues involved in the European Cooperation in Science and Technology (COST) action “Molecular Farming: Plants as production platform for high value proteins” FA0804 for helpful discussions. V.V. and T.D.M, and K.D.W. and R.P. are indebted to the “Bijzonder Onderzoeksfonds” of the Ghent University and the “Agentschap voor Innovatie door Wetenschap en Technologie” for predoctoral fellowships, respectively.
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De Buck, S. et al. (2012). Production of Camel-Like Antibodies in Plants. In: Saerens, D., Muyldermans, S. (eds) Single Domain Antibodies. Methods in Molecular Biology, vol 911. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-968-6_19
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DOI: https://doi.org/10.1007/978-1-61779-968-6_19
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