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

• Published in: Transgenic research Vol. 18; no. 5; pp. 685 - 696
• Main Authors: Joensuu, J. J, Brown, K. D, Conley, A. J, Clavijo, A, Menassa, R, Brandle, J. E
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01-10-2009; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: Animal Genetics and Genomics; Animals; Antibodies; Antibodies, Viral - biosynthesis; Antibodies, Viral - genetics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology; Coat protein; Foot-and-mouth disease; Foot-and-mouth disease virus; Foot-and-Mouth Disease Virus - immunology; Fundamental and applied biological sciences. Psychology; Genetic Engineering; Genetic technics; Immunization (passive); Immunoglobulin Variable Region - biosynthesis; Immunoglobulin Variable Region - genetics; Leaves; Life Sciences; Methods. Procedures. Technologies; Molecular Medicine; Nicotiana - genetics; Nicotiana - metabolism; Original Paper; Plant diseases; Plant Genetics and Genomics; protein purification; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Tobacco; Transgenic animals and transgenic plants; Transgenic plants; Transgenics; VP1 protein; Transgenic tobacco; Elastin-like polypeptide, ELP; Foot-and-mouth disease, FMD; Single chain variable antibody fragment, scFv; Codon optimization; Foot and mouth disease; Purification; Elastin; scFv Codon optimization; Foot-and-mouth disease FMD; Nicotiana tabacum; Fragment; Optimization; Infection; Virus; Codon; Polypeptide; Dicotyledones; Viral disease; Angiospermae; Spermatophyta; Transgenic plant; Solanaceae; Single chain variable antibody fragment; Single chain antibody
• ISSN: 0962-8819; 1573-9368
• DOI: 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.