Figure 1
From: An Effective Neutralizing Antibody Against Influenza Virus H1N1 from Human B Cells
Human monoclonal antibody production and validation. (a) Peripheral CD19 + and CD27 + memory B cells were isolated from healthy donors and immortalized by EBV. H1N1 specific clones with binding ability and neutralizing activity were screened and selected via ELISA and neutralization assays. The antibody clone 32D6 was chosen as the lead clone based on its potency. (b) 32D6 recognized H1N1, but not H5N1 and H7N9. Inactivated H1N1 (NYMC XA), H5N1 (NIBRG-14M6) and H7N9 (NIBRG-268) viruses were coated on 96-well plates. The binding intensity of serial diluted 32D6 mAbs were examined via ELISA. (c) The neutralizing activity of the 32D6 antibody against H1N1 (A/Taiwan/80813/2013), H3N2 (A/Victoria/361/2011), H5N1 (NIBRG-14M6) and H7N9 (NIBRG-268) were examined via a neutralization assay based on a high-content image system. (d) The neutralization EC50 value of 32D6 against H1N1 (A/Taiwan/80813/2013) is 0.036 μg/ml. (e) The binding kinetics of 32D6 toward HA of H1N1 was measured using the Biacore systems. The KD value is 3.528 × 10−10 M.
