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Assays for the Identification of Novel Antivirals against Bluetongue Virus
Published on: October 11, 2013
Does a SPR-Based Cell-Based Assay Provide Reliable Results on the Toxicity and Efficacy of Antiviral Drugs?
Petia Genova-Kalou1, Evdokiya Hikova2, Todor Kereziev3
1National Center of Infectious and Parasitic Diseases, 44A "Gen. Stoletov" Blvd., 1233 Sofia, Bulgaria.
Abstract:
SPR has been recently established as a powerful tool for studying various cellular processes in real time and without the use of labeling agents. So far, all studies in this area have been performed using the Kretschmann method for SPR excitation. In our studies, we used grating-based SPR. Here, we investigated the feasibility of this approach in a cell-based assay applied for antiviral drug screening. It was found that the continuous-flow SPR detection used in the conventional SPR can be replaced by sequential signal measurements of SPR slides removed from the medium at fixed hours after seeding. A protocol ensuring correct measurements was established. SPR detection was performed up to 48 h after seeding the VERO E6 cell line in three experiments, in which the cells were (i) compound-untreated, (ii) compound-treated, and (iii) infected with human coronavirus type 229E and compound-treated. Therefore, the temporal variation in the SPR signal was determined, induced by the cell coverage on the slide, the compound toxicity, and its antiviral action. MTT analysis and microscopic observations were used as reference methods. The remarkable agreement found in the results of SPR detection proved the effectiveness and reliability of grating-based SPR applied in cell-based assays.

