A New Variant of the Affinity Matrix Method for Identification of High Producing Cells in Mammalian Cell Culture

Abstract

The screening of high-producing cells is a time consuming procedure. Starting from a single cell after a single cell dilution the procedure takes weeks to complete. The affinity matrix method is a technique for screening a cell population for high producers. Using this method it is possible to perform bulk sorting via FACS and get a complete sub population containing high producing cells instead of single cells. The affinity matrix is a “molecular anchor” which captures the secreted product via a specific antibody immobilized on the cell surface. In this manner the captured product can be detected with a fluorescence labeled antibody. Cells showing a high fluorescence after this procedure have numerous products bond on their surface and are supposed to be high producers. The present work describes a variant of the affinity matrix method which can be used for antibody secreting cells. The antibodies become captured on the cell surface by immobilized protein A instead of high priced capture antibodies. We show that these affinity matrix construct is functional on the cell surface of hybridoma cells. Also we prove the bonds of the components via protein micro array and can confirm that no unspecific binding within the construct occurs.