Engineering of the membrane of fibroblast cells with virus-specific antibodies: A novel biosensor tool for virus detection

• Published in: Biosensors & bioelectronics Vol. 24; no. 4; pp. 1027 - 1030
• Main Authors: Moschopoulou, Georgia, Vitsa, Katerina, Bem, Frederic, Vassilakos, Nikos, Perdikaris, Antonis, Blouhos, Petros, Yialouris, Costas, Frosyniotis, Dimitris, Anthopoulos, Ioannis, Mangana, Olga, Nomikou, Kiki, Rodeva, Velichka, Kostova, Dimitrina, Grozeva, Stanislava, Michaelides, Alexandros, Simonian, Alex, Kintzios, Spiridon
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01-12-2008
• Subjects: Animals; Antibodies, Viral - genetics; Antibodies, Viral - immunology; Bioelectric recognition assay; Biological Assay - instrumentation; Biological Assay - methods; Biosensing Techniques - instrumentation; Biosensing Techniques - methods; Cell biosensor; Cell Membrane - physiology; Cell Membrane - virology; Cercopithecus aethiops; Cucumber green mottle mosaic virus; Cucumber mosaic virus; Cucumovirus - genetics; Cucumovirus - immunology; Cucumovirus - isolation & purification; Cytomegalovirus; Electrochemistry - instrumentation; Electrochemistry - methods; Equipment Design; Equipment Failure Analysis; Genetic Engineering - methods; Membrane-engineering; Reproducibility of Results; Sensitivity and Specificity; Tobacco rattle virus; Vero Cells; Cucumber green mottle mosaic virus; Bioelectric recognition assay; Membrane-engineering; Tobacco rattle virus; Cell biosensor
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2008.06.039

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.