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Biopharming the SimpliRED HIV diagnostic reagent in barley, potato and tobacco
- Schunmann, P.H.D., Coia, G., Waterhouse, P.M.
- Molecular breeding 2002 v.9 no.2 pp. 113-121
- transgenic plants, Hordeum vulgare, barley, Solanum tuberosum, potatoes, gene expression, recombinant antibodies, recombinant fusion proteins, epitopes, viral antigens, leaves, stems, tubers, seeds, serodiagnosis, Human immunodeficiency virus 1, hemagglutination tests
- This is the first report of an antibody-fusion protein expressed in transgenic plants for direct use in a medical diagnostic assay. By the use of gene constructs with appropriate promoters, high level expression of an anti-glycophorin single-chain antibody fused to an epitope of the HIV virus was obtained in the leaves and stems of tobacco, tubers of potato and seed of barley. This fusion protein replaces the SimpliRED diagnostic reagent, used for detecting the presence of HIV-1 antibodies in human blood. The reagent is expensive and laborious to produce by conventional means since chemical modifications to a monoclonal antibody are required. The plant-produced fusion protein was fully functional (by ELISA) in crude extracts and, for tobacco at least, could be used without further purification in the HIV agglutination assay. All three crop species produced sufficient reagent levels to be superior bioreactors to bacteria or mice, however barley grain was the most attractive bioreactor as it expressed the highest level (150 microgram of reagent g-1), is inexpensive to produce and harvest, poses a minuscule gene flow problem in the field, and the activity of the reagent is largely undiminished in stored grain. This work suggests that barley seed will be an ideal factory for the production of antibodies, diagnostic immuno-reagents, vaccines and other pharmaceutical proteins.