title: Use of virus vectors for the expression in plants of active full-length and single chain anti-coronavirus antibodies.
abstract: To extend the potential of antibodies and their derivatives to provide passive protection against enteric infections when supplied orally in crude plant extracts, we have expressed both a small immune protein (SIP) and a full-length antibody in plants using two different plant virus vectors based on potato virus X (PVX) and cowpea mosaic virus (CPMV). The alphaSIP molecule consisted of a single chain antibody (scFv) specific for the porcine coronavirus, transmissible gastroenteritis virus (TGEV) linked to the alpha-CH3 domain from human IgA. To express the full-length IgA, the individual light and heavy chains from the TGEV-specific mAb 6A.C3 were inserted into separate PVX constructs and plants were co-infected with both constructs. Western blot analysis revealed the efficient expression of both the SIP and IgA molecules. Analysis of crude plant extracts revealed that both the plant-expressed alphaSIP and IgA molecules could bind to and neutralize TGEV in tissue culture, indicating that active molecules were produced. Oral administration of crude extracts from antibody-expressing plant tissue to 2-day-old piglets showed that both the alphaSIP and full-length IgA molecules can provide in vivo protection against TGEV.
authors.affiliation: Centro Nacional de Biotecnología, Campus Universidad Autónoma, Madrid, Spain.
authors.forename: Josefa M
authors.initials: JM
authors.lastname: Alamillo
authors.forename: Wendy
authors.initials: W
authors.lastname: Monger
authors.forename: Isabel
authors.initials: I
authors.lastname: Sola
authors.forename: Beatriz
authors.initials: B
authors.lastname: García
authors.forename: Yolande
authors.initials: Y
authors.lastname: Perrin
authors.forename: Marco
authors.initials: M
authors.lastname: Bestagno
authors.forename: Oscar R
authors.initials: OR
authors.lastname: Burrone
authors.forename: Patricia
authors.initials: P
authors.lastname: Sabella
authors.forename: Joan
authors.initials: J
authors.lastname: Plana-Durán
authors.forename: Luis
authors.initials: L
authors.lastname: Enjuanes
authors.forename: George P
authors.initials: GP
authors.lastname: Lomonossoff
authors.forename: Juan A
authors.initials: JA
authors.lastname: García
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017798:Comovirus / Q000235:genetics*; D017934:Coronavirus / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D007070:Immunoglobulin A / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000276:immunology; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017863:Potexvirus / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine; D014162:Transfection / Q000379:methods
chemical_list: D000906:Antibodies; D007070:Immunoglobulin A; D007135:Immunoglobulin Variable Region; D010940:Plant Proteins; D011994:Recombinant Proteins