Purification and characterization of monoclonal IgG antibodies recognizing Ebola virus glycoprotein

Han, Baek-Sang; Jang, Ho-Young; Racine, Trina; Qiu, Xiangguo; Sin, Jeong-Im

Mice were sacrificed 3 days after the last injection and their splenocytes were prepared for fusion. The fusion was performed as previously described [9] . Briefly, the splenocytes (7×10 7 cells) were fused with 2×10 7 Sp2/0-Ag14 cells using 50% polyethylene glycol 4000 (PEG4000). The cells were incubated at 37°C for 10 days in cDMEM containing hypoxanthine-aminopterin-thymidine (HAT). Subsequently, the cells were incubated with cDMEM containing hypoxanthine-thymidine (HT). After 15 days, the cell supernatants were screened for Ab-secreting hybridomas using enzyme-linked immunosorbent assay (ELISA). For production of MAbs, the Ab-secreting hybridomas were diluted to make less than a single cell per well using the limiting dilution sub-culture method [11] . PEG4000, HAT and HT were purchased from Sigma-Aldrich (St. Louis, MO, USA).