Hybridoma screening for cell adhesion molecules using multiple parallel comparisons in conditions of flow

Cell adhesion is a complex biophysical process that plays a central role in immunophysiology. Because of the complex force-energy relationships involved, insights into the mechanism of cell adhesion largely depend on comparative measurements. In this report, we describe a comparative approach to the measurement of cell adhesion under conditions of flow. The assay system perfuses fluorescently labeled lymphocytes over a cell monolayer in commercially available multiwell culture plates. The fluorescently labeled cells demonstrate a reproducible flow pattern within the well. Videomicroscopic recordings of cell movement have demonstrated rolling behavior over a wide range of cell velocities. This technique permits the measurement of cell adhesion over a variety of flow velocities, time courses, and treatment conditions. The ability to vary treatment conditions and provide multiple parallel conditions suggests the utility of this approach in the development of monoclonal antibodies (MAb) recognizing cell adhesion molecules.