Expression and purification of an anti-Foot-and-mouth disease virus single chain variable antibody fragment in tobacco plants
Joensuu, J.J., Brown, K.D., Conley, A.J., Clavijo, A., Menassa, R., Brandle, J.E. (2009). Expression and purification of an anti-Foot-and-mouth disease virus single chain variable antibody fragment in tobacco plants, 18(5), 685-696. http://dx.doi.org/10.1007/s11248-009-9257-0
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. © Her Majesty the Queen in Right of Canada 2009.
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