Biopharming the SimpliRED™ HIV diagnostic reagent in barley, potato and tobacco.

Autor: Schünmann, Petra, Coia, Greg, Waterhouse, Peter
Zdroj: Molecular Breeding; Jun2002, Vol. 9 Issue 2, p113-121, 9p
Abstrakt: This is the first report of an antibody-fusion protein expressed intransgenic plants for direct use in a medical diagnostic assay. By the use ofgene constructs with appropriate promoters, high level expression of ananti-glycophorin single-chain antibody fused to an epitope of the HIV virus wasobtained in the leaves and stems of tobacco, tubers of potato and seed ofbarley. This fusion protein replaces the SimpliRED™ diagnostic reagent,used for detecting the presence of HIV-1 antibodies in human blood. The reagentis expensive and laborious to produce by conventional means since chemicalmodifications to a monoclonal antibody are required. The plant-produced fusionprotein was fully functional (by ELISA) in crude extracts and, for tobacco atleast, could be used without further purification in the HIV agglutinationassay. All three crop species produced sufficient reagent levels to be superiorbioreactors to bacteria or mice, however barley grain was the most attractivebioreactor as it expressed the highest level (150 μg of reagentg-1), is inexpensive to produce and harvest, poses aminuscule gene flow problem in the field, and the activity of the reagent islargely undiminished in stored grain. This work suggests that barley seed willbe an ideal factory for the production of antibodies, diagnosticimmuno-reagents, vaccines and other pharmaceutical proteins. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index