title: In vivo expression and binding activity of scFv-RWAV, which recognizes the coat protein of tomato leaf curl New Delhi virus (family Geminiviridae).
abstract: Recombinant antibodies expressed in plants have the potential to interrupt virus infections by blocking essential stages of the infection cycle. Here, we show that the expression of a recombinant single-chain variable fragment (scFv) that recognizes the coat protein of tomato leaf curl New Delhi virus (ToLCNDV) in vitro can also bind to a recombinant coat protein in vivo in the reducing environment of the plant cytosol. The scFv and its target were both expressed as fluorescent protein fusions, one incorporating green fluorescent protein (GFP) and the other DsRed. We found that the incorporation of a nuclear localization signal into the scFv construct resulted in the nuclear import of the antibody-antigen complex, as shown by colocalization of the two fluorescent signals. This demonstrates that recombinant antibodies can be targeted to the nucleus and will bind to geminivirus coat proteins therein, allowing the virus infection cycle to be interrupted during its critical replicative phase.
authors.affiliation: Institute for Molecular Biotechnology (Biology VII), RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany. adelzakri@gmail.com
authors.forename: Adel M
authors.initials: AM
authors.lastname: Zakri
authors.forename: Angelika
authors.initials: A
authors.lastname: Ziegler
authors.forename: Ulrich
authors.initials: U
authors.lastname: Commandeur
authors.forename: Rainer
authors.initials: R
authors.lastname: Fischer
authors.forename: Lesley
authors.initials: L
authors.lastname: Torrance
mesh_terms: D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D017791:Geminiviridae / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D018613:Microscopy, Confocal; D010935:Plant Diseases / Q000821:virology; D011485:Protein Binding / Q000502:physiology; D014026:Nicotiana / Q000821:virology*
chemical_list: D036022:Capsid Proteins