Biologia plantarum 52:184-186, 2008 | DOI: 10.1007/s10535-008-0041-4
Optimum storage conditions for product of transiently expressed epitopes of Human papillomavirus using Potato virus X-based vector
- 1 Institute of Experimental Botany, Academy of Sciences of Czech Republic, Praha, Czech Republic
We describe the optimized storage conditions of recombinant Potato virus A coat protein (ACP) carrying two different epitopes from Human papillomavirus type 16 (HPV-16). Epitope derived from minor capsid protein L2 was expressed as N-terminal fusion with ACP while an epitope derived from E7 oncoprotein was fused to its C-termini. The construct was cloned into Potato X potexvirus (PVX) based vector and transiently expressed in plants using Agrobacterium tumefaciens mediated inoculation. The effect of storage conditions on the serological activity of L2ACPE7 was studied by ELISA using IgG anti PVX, PVA and L2. Purified L2ACPE7 stored freeze-dried (at -20 °C), frozen at various temperatures (-20 °C, -70 °C) and at +4 °C were tested. Purified L2ACPE7 was most stable as lyophilized material stored at -20 °C. Our study demonstrates suitable way for the storage of plant material containing foreign viral epitopes for the purposes of edible vaccination.
Keywords: edible vaccination; heterologous proteins; Nicotiana benthamiana; serological activity
Subjects: Nicotiana benthamiana; polyclonal antibodies; Potato virus; proteins; viral epitopes
Received: February 16, 2007; Accepted: August 28, 2007; Published: March 1, 2008 Show citation
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