Author: Laura Riva; Shuofeng Yuan; Xin Yin; Laura Martin-Sancho; Naoko Matsunaga; Sebastian Burgstaller-Muehlbacher; Lars Pache; Paul P. De Jesus; Mitchell V. Hull; Max Chang; Jasper Fuk-Woo Chan; Jianli Cao; Vincent Kwok-Man Poon; Kristina Herbert; Tu-Trinh Nguyen; Yuan Pu; Courtney Nguyen; Andrey Rubanov; Luis Martinez-Sobrido; Wen-Chun Liu; Lisa Miorin; Kris M. White; Jeffrey R. Johnson; Christopher Benner; Ren Sun; Peter G. Schultz; Andrew Su; Adolfo Garcia-Sastre; Arnab K. Chatterjee; Kwok-Yung Yuen; Sumit K. Chanda
Title: A Large-scale Drug Repositioning Survey for SARS-CoV-2 Antivirals Document date: 2020_4_17
ID: 1fgnfh62_19
Snippet: The selected hits were further validated by immunofluorescence in an 8-point dose response experiment to determine EC 50 After blocking with 3 % bovine serum albumin (BSA) for 30 min, the cells were incubated for 1 hour at room temperature with rabbit-anti-SARS-CoV-1 nucleoprotein serum, which exhibits strong cross-reactivity with SARS-CoV-2. After two washes with phosphate-buffered saline (PBS), the cells were incubated with Alexa Fluor 488conju.....
Document: The selected hits were further validated by immunofluorescence in an 8-point dose response experiment to determine EC 50 After blocking with 3 % bovine serum albumin (BSA) for 30 min, the cells were incubated for 1 hour at room temperature with rabbit-anti-SARS-CoV-1 nucleoprotein serum, which exhibits strong cross-reactivity with SARS-CoV-2. After two washes with phosphate-buffered saline (PBS), the cells were incubated with Alexa Fluor 488conjugated goat-anti-rabbit IgG (Thermo Fisher Scientific, USA) for 1 hour at room temperature. After two additional washes, PBS supplemented with 0.1 µg/ml antifade-4 6-diamidino-2-phenylindole (DAPI) (BioLegend, USA) was added to the cells for at least 30 min before imaging. Images were acquired using the Celigo Image Cytometer (Nexcelom). The assay results and data analysis enabled us to determine infectivity and viability/cytotoxicity. Based on all infectivity and cytotoxicity values, a 4-parameter logistic non-linear regression model was used to calculate EC 50 and CC 50 concentration values.
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