The production of a genus-specific recombinant antibody (scFv) using a recombinant potyvirus protease.
A single chain variable fragment antibody (scFv; anti-NIa scFv102) was selected from a synthetic human antibody library by using a NIa protease of Plum pox virus (PPV) as an antigen, which was expressed in bacteria. The NIa protease forms the nuclear inclusion body A and acts as the major protease in the cleavage of the viral polyprotein into functional proteins. The NIa protein was detected with anti-NIa scFv102 after expression in Escherichia coli cells as well as from PPV-infected Nicotiana benthamiana plants. Furthermore, the scFv102 has the ability to identify not only PPV from infected plants but also can detect other infections with members of the potyviruses. Nineteen different potyviruses were recognized by the scFv102 in various infected plants tested through dot blot assays. Therefore, the antibody scFv102 has the potential of becoming a general tool to detect potyvirus infections in different plant species.
Hust
Michael
M
Lehrgebiet Molekulargenetik, Universität Hannover, Herrenhäuser Strasse 2, D-30419 Hannover, Germany.
Maiss
Edgar
E
Jacobsen
Hans-Jörg
HJ
Reinard
Thomas
T
D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000276:immunology; D004926:Escherichia coli; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D017801:Plum Pox Virus / Q000201:enzymology / Q000276:immunology*; D017800:Potyvirus / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013045:Species Specificity; D014026:Nicotiana / Q000821:virology; D014764:Viral Proteins / Q000276:immunology
D000914:Antibodies, Viral; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D014764:Viral Proteins; D010450:Endopeptidases; C059992:nuclear inclusion protein a, mosaic viruses