Surface plasmon resonance for monitoring the interaction of Potato virus Y with monoclonal antibodies.
Surface plasmon resonance (SPR)-based biosensors have been widely utilized for measuring interactions of a variety of molecules. Fewer examples include higher biological entities such as bacteria and viruses, and even fewer deal with plant viruses. Here, we describe the optimization of an SPR sensor chip for evaluation of the interaction of the economically relevant filamentous Potato virus Y (PVY) with monoclonal antibodies. Different virus isolates were efficiently and stably bound to a previously immobilized polyclonal antibody surface, which remained stable over subsequent injection regeneration steps. The ability of the biosensor to detect and quantify PVY particles was compared with ELISA and RT-qPCR. Stably captured virus surfaces were successfully used to explore kinetic parameters of the interaction of a panel of monoclonal antibodies with two PVY isolates representing the main viral serotypes N and O. In addition, the optimized biosensor proved to be suitable for evaluating whether two given monoclonal antibodies compete for the same epitope within the viral particle surface. The strategy proposed in this work can help to improve existing serologic diagnostic tools that target PVY and will allow investigation of the inherent serological variability of the virus and exploration for new interactions of PVY particles with other proteins.
Department of Biotechnology and Systems Biology, National Institute of Biology, 1000 Ljubljana, Slovenia; Centre of Excellence for Biosensors, Instrumentation, and Process Control-COBIK, Solkan, Slovenia. Electronic address: ion.gutierrez@nib.si.
Ion
I
Gutiérrez-Aguirre
Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia.
Vesna
V
Hodnik
INRA, UMR 1349 IGEPP, F-35653 Le Rheu, France; FN3PT/RD3PT, 75008 Paris, France.
Laurent
L
Glais
Department of Biotechnology and Systems Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.
Matevž
M
Rupar
INRA, UMR 385 BGPI, Campus de Baillarguet, 34398 Montpellier, France.
Emmanuel
E
Jacquot
Department of Biology, Biotechnical Faculty, University of Ljubljana, 1000 Ljubljana, Slovenia; Laboratory for Molecular Biology and Nanobiotechnology, National Institute of Chemistry, 1000 Ljubljana, Slovenia; Centre of Excellence En-Fist, 1000 Ljubljana, Slovenia. Electronic address: gregor.anderluh@ki.si.
Gregor
G
Anderluh
Department of Biotechnology and Systems Biology, National Institute of Biology, 1000 Ljubljana, Slovenia; Centre of Excellence for Biosensors, Instrumentation, and Process Control-COBIK, Solkan, Slovenia.
Maja
M
Ravnikar
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology; D017800:Potyvirus / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance / Q000379:methods*
D000911:Antibodies, Monoclonal; D000939:Epitopes
Biosensor; Filamentous virus; Monoclonal antibody; PVY; Surface plasmon resonance