Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications

Mohammadzadeh, Sara; Khabiri, Alireza; Roohvand, Farzin; Memarnejadian, Arash; Salmanian, Ali Hatef; Ajdary, Soheila; Ehsani, Parastoo

ELISA was performed using the streptavidin-biotin peroxidase complex (ABC) assay. Briefly, wells were coated overnight at room temperature with 100 µL of 5 µg/mL mouse monoclonal antibody against 21-40 amino acids of HCV core antigen (Abcam, UK) and blocked with 1% bovine serum albumin (BSA) for two hours at 37˚C. After several washing steps, 50 μg of TSP from each sample or eHCVcp (for standard curve, serial dilutions in the range of 0.1-1 µg/mL) were added to the coated wells in duplicates and incubated for one hour at 37°C. Following the washing steps, biotinylated anti-core polyclonal antibodies (1:1000 dilution, Abcam, UK) were immersed into the wells and incubation continued at 37˚C for one hour followed by washing steps and another similar incubation with streptavidin HRP conjugate (1:4000; Sigma). Finally, by adding TMB and color development, the absorbance at 450 nm was measured.