) This is a new facility to be located in recently renovated space. For several years, one laboratory (Dr. Lees) has served as a resource helping other groups to generate monoclonals. Monoclonal antibodies are a fundamental tool in establishing the precise role of proteins in vivo. The need to provide this as a shared facility is clear. A central facility will ensure maintenance of well tested and reliable myeloma cell lines, serum and other reagents that will greatly improve efficiency and cost effectiveness of the fusions. This will also allow production of antibodies that would otherwise not be generated because of lack of expertise and, in that way, significantly enhance the progress of the research. The facility will operate as a near full service unit using the standard operating protocol defined by the Lees lab. It will conduct most of the steps of monoclonal production including- (1) injection and bleeding of mice, (2) hybridoma fusions, (3) primary ELISA screening, (4) expansion and freezing of primary positive clones (up to 50 per fusion), (5) single cell cloning (up to 10 lines per fusion), (6) expansion and freezing of selected lines (10 vials per line) and (7) large scale production of tissue culture supernatants. Professor Jacqueline Lees will be the facility supervisor. All consumables will be covered by chargeback fees. We propose to charge user fees to cover the cost of purchase, housing, immunization and bleeding of mice 2 P30 CA14051-29 29 Dr. Richard O. Hynes ($250 per antigen) and the cost of the hybridoma fusion and antibody production ($1500 per fusion).
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