Simultaneous delivery of an active protein and neutralizing antibody: creation of separated regions of biological activity.

Spatial control over the biological activity of nerve growth factor (NGF) via a novel type of controlled-release device was demonstrated in an in vitro system. Two-layer polymer matrices that simultaneously released NGF and a neutralizing antibody (anti-NGF) from opposite faces were placed in PC12 cell-populated collagen gels. Biological activity in the gels was assessed over the course of 10 days by direct observation of the cells, which extend neuronal processes in the presence of NGF in a dose-dependent manner. The concentrations of both proteins in the gels were determined by ELISA as a function of distance from the polymer matrices at various time points. A boundary in biological activity was established within a few days of the initiation of the cultures; this boundary persisted and became more pronounced throughout the duration of the experiment. ELISA analysis revealed regions of high concentration of both NGF and anti-NGF on their respective sides of the polymer matrix early in the experiment. The theoretical amount of active NGF in the gel sections was calculated on the basis of these ELISA results; the concentration of active NGF in the region adjacent to the polymer correlated with the observed degree of biological response. These experiments suggest that spatial control over the biological activity of a potent agent can be obtained by an appropriately designed controlled-release device.