Quick antibody adjuvant

MAbs were produced as described previously [28 (link)]. Briefly, 6-week-old, female, BALB/c mice (Shanghai Laboratory Animal Center) were immunized with two intramuscular injection of H7N9 virus vaccine in Quick Antibody adjuvant (Biodragon, China) with 2-week interval. The mice then received an additional intravenous injection of the same viral antigen 3 days without adjuvant before the fusion of splenocytes with SP2/0 myeloma cells [29 (link)]. After cell fusion, hybridoma culture supernatants were screened by enzyme-linked immunosorbent assay (ELISA) [30 (link)]. Through limiting dilution, positive hybridoma lines were cloned, expanded and further sub-cultured. Hybridoma cell clones were expanded significantly, harvested, and injected intraperitoneally in pristane-primed BALB/c mice [29 (link)]. The ascites was collected and purified by protein-G column (GE) according to the manufacturer’s instructions. The concentrations of purified mAbs were determined by Nanodrop 2000 (Thermo), and were stored at −80°C. The subclass and type of mAbs were determined by using the Mouse monoclonal antibody isotyping kit (Bio-Rad) as directed by the manufacturer’s instructions. The heavy and light chains of the hybridomas are sequenced by Genscript (Nanjing, China).

MAbs were produced as described previously [28 (link)]. Briefly, 6-week-old, female, BALB/c mice (Shanghai Laboratory Animal Center) were immunized with two intramuscular injection of H7N9 virus vaccine in Quick Antibody adjuvant (Biodragon, China) with 2-week interval. The mice then received an additional intravenous injection of the same viral antigen 3 days without adjuvant before the fusion of splenocytes with SP2/0 myeloma cells [29 (link)]. After cell fusion, hybridoma culture supernatants were screened by enzyme-linked immunosorbent assay (ELISA) [30 (link)]. Through limiting dilution, positive hybridoma lines were cloned, expanded and further sub-cultured. Hybridoma cell clones were expanded significantly, harvested, and injected intraperitoneally in pristane-primed BALB/c mice [29 (link)]. The ascites was collected and purified by protein-G column (GE) according to the manufacturer’s instructions. The concentrations of purified mAbs were determined by Nanodrop 2000 (Thermo), and were stored at −80°C. The subclass and type of mAbs were determined by using the Mouse monoclonal antibody isotyping kit (Bio-Rad) as directed by the manufacturer’s instructions. The heavy and light chains of the hybridomas are sequenced by Genscript (Nanjing, China).