Use of virus vectors for the expression in plants of active full-length and single chain anti-coronavirus antibodies.
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.
Alamillo
Josefa M
JM
Centro Nacional de Biotecnología, Campus Universidad Autónoma, Madrid, Spain.
Monger
Wendy
W
Sola
Isabel
I
García
Beatriz
B
Perrin
Yolande
Y
Bestagno
Marco
M
Burrone
Oscar R
OR
Sabella
Patricia
P
Plana-Durán
Joan
J
Enjuanes
Luis
L
Lomonossoff
George P
GP
García
Juan A
JA
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
D000906:Antibodies; D007070:Immunoglobulin A; D007135:Immunoglobulin Variable Region; D010940:Plant Proteins; D011994:Recombinant Proteins