Fig. 4
From: Production of antigen-specific human IgGs by in vitro immunization
An optimal anti-Tat specific B-lymphocyte response was reached after 11 days of in vitro immunization. PBMCs were incubated for eight, eleven or thirteen days in the presence or absence of anti-CD40/IL-4/IL-21, ZZTat101 + anti-CD40/IL-4/IL-21, respectively. Then, PBMCs were transferred from culture plates to ELISPOT plates previously coated with Tat101. After an additional 24-h incubation at 37 °C, a biotinylated anti-human IgG was added. Spot-forming cells were then detected using a streptavidin-alkaline phosphatase conjugate and NBT/BCIP as substrate. Spot-forming cells were quantified with an automated ELISPOT reader