Title

High-yield expression of a viral peptide animal vaccine in transgenic tobacco chloroplasts

Authors

Authors

A. Molina; S. Hervas-Stubbs; H. Daniell; A. M. Mingo-Castel;J. Veramendi

Abbreviated Journal Title

Plant Biotechnol. J.

Keywords

canine parvovinus; CTB; fusion protein; GFP; peptide vaccine; plastid; transformation; TOXIN-B-SUBUNIT; NSP4 FUSION PROTEIN; CHOLERA-TOXIN; CANINE PARVOVIRUS; EDIBLE VACCINES; GENE-EXPRESSION; TRANSLATION EFFICIENCY; SYNTHETIC; PEPTIDE; SURFACE-ANTIGEN; DISEASE VIRUS; Biotechnology & Applied Microbiology; Plant Sciences

Abstract

The 2L21 peptide, which confers protection to dogs against challenge with virulent canine parvovirus (CPV), was expressed in tobacco chloroplasts as a C-terminal translational fusion with the cholera toxin B subunit (CTB) or the green fluorescent protein (GFP). Expression of recombinant proteins was dependent on plant age. A very high-yield production was achieved in mature plants at the time of full flowering (310 mg CTB-2L21 protein per plant). Both young and senescent plants accumulated lower amounts of recombinant proteins than mature plants. This shows the importance of the time of harvest when scaling up the process. The maximum level of CTB-2L21 was 7.49 mg/g fresh weight (equivalent to 31.1% of total soluble protein, TSP) and that of GFP-2L21 was 5.96 mg/g fresh weight (equivalent to 22.6% of TSP). The 2L21 inserted epitope could be detected with a CPV-neutralizing monoclonal antibody, indicating that the epitope is correctly presented at the C-terminus of the fusion proteins. The resulting chimera CTB-2L21 protein retained pentamerization and G(M1)-ganglioside binding characteristics of the native CTB and induced antibodies able to recognize VP2 protein from CPV. To our knowledge, this is the first report of an animal vaccine epitope expression in transgenic chloroplasts. The high expression of antigens in chloroplasts would reduce the amount of plant material required for vaccination (similar to100 mg for a dose of 500 mug antigen) and would permit encapsulation of freeze-dried material or pill formation.

Journal Title

Plant Biotechnology Journal

Volume

2

Issue/Number

2

Publication Date

1-1-2004

Document Type

Article

Language

English

First Page

141

Last Page

153

WOS Identifier

WOS:000220101600005

ISSN

1467-7644

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