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Expression in tobacco and purification of beak and feather disease virus capsid protein fused to elastin-like polypeptides

Author:
Duvenage, Lucian, Hitzeroth, Inga I., Meyers, Ann E., Rybicki, Edward P.
Source:
Journal of virological methods 2013 v.191 no.1 pp. 55-62
ISSN:
0166-0934
Subject:
Beak and feather disease virus, Nicotiana benthamiana, beak, coat proteins, feathers, polypeptides, subunit vaccines, tobacco
Abstract:
Psittacine beak and feather disease, caused by beak and feather disease virus (BFDV), is a threat to endangered psittacine species. There is currently no vaccine against BFDV, which necessitates the development of safe and affordable vaccine candidates. A subunit vaccine based on BFDV capsid protein (CP), the major antigenic determinant, expressed in the inexpensive and highly scalable plant expression system could satisfy these requirements. Full-length CP and a truncated CP (ΔN40 CP) were transiently expressed in tobacco (Nicotiana benthamiana) as fusions to elastin-like polypeptide (ELP). These two proteins were fused to ELPs of different lengths in order to increase expression levels and to provide a simple means of purification. The ELP fusion proteins were purified by inverse transition cycling (ITC) and it was found that a membrane filtration-based ITC method improved the recovery of ΔN40 CP-ELP51 fusion protein relative to a centrifugation-based method.
Agid:
877747