Lymphocyte Culture Core

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The Lymphocyte Culture Core (LCC), a hybridoma facility, is a research support facility of the University of Virginia School of Medicine and the Cancer Center. The primary function of the LCC is to make available to researchers the most current technology and expertise for the construction and selection of lymphocytemyeloma hybridomas for the production of monoclonal antibodies. The LCC also provides expertise in the use of these reagents in basic research programs. Services are customized to the specific requirements of individual investigators to optimize the recovery of appropriate antigen specific monoclonal antibodies. Initial discussions are held with the investigator and his staff in order to understand the experimental system employed in the laboratory and to determine the specific applications for which monoclonal antibodies are required. The staff of the LCC then plans and executes all aspects of hybridoma construction and selection and monoclonal antibody production. Specifically, the LCC provides approved animal experimentation protocols for all aspects of monoclonal antibody production, including: immunizations, sera collection, intrasplenic injection and ascites production. Center staff immunize animals, collect sera, perform all cell culture aspects of hybridoma construction and selection, and develop ELISA assay strategies appropriate for sera titration and for screening large numbers of hybridoma culture supernatants. Once an appropriate assay system has been developed and used to demonstrate the successful immunization of the necessary animals, fusions are scheduled on a first come first served basis. The center conducts all aspects of cell fusion, assaying by ELISA for specific antibody positive cultures, culturing, cloning, freezing, and recovery of specific antibody producing clones. Investigators receive, upon request, hybridoma culture supernatants and individual hybridoma clones. The center also provides cryopreservation, storage and recovery of cryopreserved cells. Adjunct services include monoclonal antibody isotyping and sub-isotyping, bulk monoclonal antibody production in vitro using disposable bioreactors, ascites production, monoclonal and polyclonal antibody purification by affinity chromatography on recombinant Protein G columns, and antibody inventory, storage and shipping. The LCC also serves as a repository, production facility and distribution point for intellectual property (hybridomas and monoclonal antibodies) licensed by the University of Virginia Patent Foundation.