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Study on the mechanism of Bioelectric Recognition Assay: evidence for immobilized cell membrane interactions with viral fragments

病毒 生物 细胞 生物传感器 生物物理学 细胞生物学 细胞培养 蛋白酶 化学 生物化学 病毒学 遗传学
作者
Spyridon Kintzios,F. Bem,Olga Mangana,Kyriaki Nomikou,Panayotis Markoulatos,N. Alexandropoulos,Costas Fasseas,V.B. Arakelyan,Andreas Petrou,Katerina Soukouli,Georgia Moschopoulou,Constantine P. Yialouris,Aleksandr Simonian
出处
期刊:Biosensors and Bioelectronics [Elsevier BV]
卷期号:20 (4): 907-916 被引量:25
标识
DOI:10.1016/j.bios.2004.04.009
摘要

The Bioelectric Recognition Assay (BERA) is a whole-cell based biosensing system that detects the electric response of cultured cells, suspended in a gel matrix, to various ligands, which bind to the cell and/or affect its physiology. Previous studies have demonstrated the potential application of this method for rapid, inexpensive detection of viruses in a crude sample. However, the understanding, so far, of the fundamental processes that take place during cell–virus interactions within the probe has been rather limited. In the present study, we combined electrophysiological and fluorescence microscopical assays, so that we can prove that animal and plant cells immobilized in BERA sensors respond to different viruses primarily by changing their membrane potential. The response of immobilized cells against different viruses did not depend on the virus ability to penetrate the cell, but was modified after binding each virus to a virus-specific antibody or removal of its coat protein after treatment with a protease. Consequently, we were able to assay the presence of a virus in its complete form or fragments thereof. Combination of immunological recognition with the electrophysiological response of immobilized cells allows for a considerable increase of the specificity of the BERA biosensory assay. In addition, rather than simply detect the presence of a protein or genomic sequence, the method can help gain information on the bioactivity of a virus.
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