Fig. 6
The generated HCV4a-VLPs exhibited physical properties comparable to HCV native virus. HCV4a-HEK293T stable cells were cultured and harvested for the preparation of cell lysate. The HCV4a-VLPs were isolated and purified from the cleared lysate by the ultracentrifugation on 30% sucrose cushion for 2 hours at 40,000 rpm at 4oC. The isolated HCV4a-VLPs were resuspended in PBS and examined by ELISA and TEM assays. a An ELISA assay for detecting the presence of HCV4a-VLPs in the pellet was conducted by detection of HCV core protein in the purified VLPs. The ELISA results confirmed the enrichment of the HCV core in the virus pellet after the isolation of VLPs by ultracentrifugation suggesting the successful formation and isolation of HCV4a-VLPs. The data are represented as mean ± SD of three replicates b The purified VLPs were investigated by the negative-staining TEM analysis. The results of TEM analysis revealed the presence of spherical VLPs particles of 60–65 nm average size similar to the shape and average size of the previously reported HCV native virus