PROVENGE® consists of autologous peripheral blood mononuclear cells, including antigen presenting cells (APCs), that have been activated, during a defined culture period, with a recombinant human protein, PAP-GM-CSF, consisting of prostatic acid phosphatase (PAP), an antigen expressed in prostate cancer tissue, linked to granulocyte-macrophage colony stimulating factor (GM-CSF), an immune cell activator.
Dendritic-cell precursors are harvested from the peripheral blood by a standard 1.5-2.0 blood volume mononuclear cell leukapheresis. The leukapheresis product is layered over a buoyant density solution (1.077 g/mL) and centrifuged at 1,000 g for 20 minutes to deplete erythrocytes and granulocytes. The interface cells are collected, washed, layered over a second buoyant density solution (1.065 g/mL), and centrifuged at 805 g for 30 minutes to deplete platelets and low-density monocytes and lymphocytes. The cell pellet, which contains dendritic-cell precursors, is washed and incubated in AIM media with PA2024 (10 μg/mL). The culture medium does not contain serum or exogenous cytokines. After incubation for 40 hours at 37°C in 5% CO2 atmosphere, the cells are washed and formulated at the desired clinical dose in 250 mL of lactated Ringers’ solution.