title: Engineering of the membrane of fibroblast cells with virus-specific antibodies: A novel biosensor tool for virus detection.
abstract: A novel concept for the assay of viral antigens is described. The methodological approach is based on a membrane-engineering process involving the electroinsertion of virus-specific antibodies in the membranes of fibroblast cells. As a representative example, Vero fibroblasts were engineered with antibodies against Cucumber mosaic virus (CMV) and used for the construction of an ultra-sensitive miniature cell biosensor system. The attachment of a homologous virus triggered specific changes to the cell membrane potential that were measured by appropriate microelectrodes, according to the principle of the bioelectric recognition assay (BERA). No change in the membrane potential was observed upon cell contact with the heterologous cucumber green mottle mosaic virus (CGMMV). Fluorescence microscopy observations showed that attachment of CMV particles to membrane-engineered cells was associated with membrane hyperpolarization and increased [Ca(2+)](cyt). In an additional field-based application, we were able to detect CMV-infected tobacco plants at an essentially 100% level of accuracy.
authors.affiliation: Laboratory of Plant Physiology, Faculty of Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece; EMBIO Diagnostics, Nicosia, Cyprus.
authors.forename: Georgia
authors.initials: G
authors.lastname: Moschopoulou
authors.forename: Katerina
authors.initials: K
authors.lastname: Vitsa
authors.forename: Frederic
authors.initials: F
authors.lastname: Bem
authors.forename: Nikos
authors.initials: N
authors.lastname: Vassilakos
authors.forename: Antonis
authors.initials: A
authors.lastname: Perdikaris
authors.forename: Petros
authors.initials: P
authors.lastname: Blouhos
authors.forename: Costas
authors.initials: C
authors.lastname: Yialouris
authors.forename: Dimitris
authors.initials: D
authors.lastname: Frosyniotis
authors.forename: Ioannis
authors.initials: I
authors.lastname: Anthopoulos
authors.forename: Olga
authors.initials: O
authors.lastname: Mangana
authors.forename: Kiki
authors.initials: K
authors.lastname: Nomikou
authors.forename: Velichka
authors.initials: V
authors.lastname: Rodeva
authors.forename: Dimitrina
authors.initials: D
authors.lastname: Kostova
authors.forename: Stanislava
authors.initials: S
authors.lastname: Grozeva
authors.forename: Alexandros
authors.initials: A
authors.lastname: Michaelides
authors.forename: Alex
authors.initials: A
authors.lastname: Simonian
authors.forename: Spiridon
authors.initials: S
authors.lastname: Kintzios
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002462:Cell Membrane / Q000502:physiology* / Q000821:virology*; D002522:Chlorocebus aethiops; D017799:Cucumovirus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005818:Genetic Engineering / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014709:Vero Cells
chemical_list: D000914:Antibodies, Viral