Rapid discovery and optimization of therapeutic antibodies against emerging infectious diseases

Rogers, J.; Schoepp, R.J.; Schröder, O.; Clements, T.L.; Holland, T.F.; Li, J.Q.; Li, J.; Lewis, L.M.; Dirmeier, R.P.; Frey, G.J.; Tan, X.; Wong, K.; Woodnutt, G.; Keller, M.; Reed, D.S.; Kimmel, B.E.; Tozer, E.C.

Ten Balb/c mice were immunized with sucrose gradient purified SARS virus, Centers for Disease Control and Prevention (CDC) strain 809921 (originally provided by Dr Thomas G. Ksiazek, Special Pathogens Branch, CDC, Atlanta, GA, USA) . Each mouse was initially dosed with 1 Â 10 4 plaque forming units ( pfu) of live virus by the intraperitoneal (i.p.) route on days 0, 14 and 28; followed by the same dose of g-irradiated virus on day 47. Antibody titers on a SARS-CoV ELISA ranged from 1:25 600 to 1:409 600. Spleens of three mice were removed, processed through a sterile mesh screen, washed with Dulbecco's minimal essential medium (DMEM) with penicillin (200 IU/ml) and streptomycin (200 mg/ml), and pelleted by centrifugation for 5 min at 1000 r.p.m. The cell pellet was resuspended in DMEM containing antibiotics, 10% DMSO, and fetal bovine serum and stored in liquid nitrogen.