Engineering of the membrane of fibroblast cells with virus-specific antibodies: A novel biosensor tool for virus detection.
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.
Moschopoulou
Georgia
G
Laboratory of Plant Physiology, Faculty of Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece; EMBIO Diagnostics, Nicosia, Cyprus.
Vitsa
Katerina
K
Bem
Frederic
F
Vassilakos
Nikos
N
Perdikaris
Antonis
A
Blouhos
Petros
P
Yialouris
Costas
C
Frosyniotis
Dimitris
D
Anthopoulos
Ioannis
I
Mangana
Olga
O
Nomikou
Kiki
K
Rodeva
Velichka
V
Kostova
Dimitrina
D
Grozeva
Stanislava
S
Michaelides
Alexandros
A
Simonian
Alex
A
Kintzios
Spiridon
S
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
D000914:Antibodies, Viral