CORE--LYMPHOCYTE/HYBRIDOMA

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Description: (Applicant's Description) The Lymphocyte Culture/Hybridoma Center (LCC) 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 lymphocyte-myeloma 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 an investigator and his staff in order to understand the experimental system employed in his laboratory and to determine the specific applications for which monoclonal antibodies are required. The staff of the LCC then plan and execute all aspects of hybridoma construction and selection and monoclonal antibody production. LCC staff immunize animals, collect sera and develop ELISA assay strategies appropriate for screening large numbers of hybridoma culture supernatants and for the titration of animal sera. 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 the individual hybridoma clones. The center also provides other extended services using bulk expansion of clones, large scale in vitro production of monoclonal antibodies using gas permeable bags and hollow fiber bioreactor technology, ascites antibody production, isotyping and subisotyping of monoclonal antibodies, long term cryogenic storage of cells, and the routine analytical and preparative purification of monoclonal and polyclonal antibodies by affinity chromatography on recombinant Protein G columns.