Čeština
EnglishOptimum storage conditions for product of transiently expressed epitopes of Human papillomavirus using Potato virus X-based vector
Čeřovská N., Hoffmeisterová H., Moravec T., Plchová H., Folwarczna J., Hadámková R.
BIOLOGIA PLANTARUM 52: 184-186, 2008
Keywords: edible vaccination, heterologous proteins, Nicotiana benthamiana, serological activity
Abstract: 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.
Fulltext: contact IEB authors
IEB authors: Jitka Folwarczna, Hana Hoffmeisterová, Tomáš Moravec, Helena Plchová
BIOLOGIA PLANTARUM 52: 184-186, 2008
Keywords: edible vaccination, heterologous proteins, Nicotiana benthamiana, serological activity
Abstract: 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.
Fulltext: contact IEB authors
IEB authors: Jitka Folwarczna, Hana Hoffmeisterová, Tomáš Moravec, Helena Plchová