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SummaryThe Immunology Core's primary service offered is the production of mouse monoclonal antibodies for UNC investigators. Alternatively, the core can produce hamster and rat monoclonal antibodies. These services are generally provided on a first-come, first-served basis. Successful hybridoma production depends on a several factors that need to be adequately addressed before a monoclonal project is initiated. The most important factors to the customers who desire quality monoclonal reagents are the purity, quantity and immunogenicity of the antigen used for immunization and the screening strategy that will be employed to quickly identify the antibody-secreting hybridomas of interest. The antigen used in the immunization of the mice should be as pure as possible. About 1-4 mg of soluble antigen will be needed for immunization and the screening assays*. It is necessary to screen the clones on pure material, so if pure material is limited, crude material can be used for the immunizations. If peptide immunogens are used, it is important to select the peptide after considering the following predictors of antigenicity: hydrophilicity, amphipathicity, surface accessibility, chain termination and sequence variability. The core can provide limited assistance into the selection of peptides based on these predictors. The Immunology Core can do the conjugation of peptides to carrier proteins in order to enhance their immunogenicity. The screening strategy needs to be discussed with the core prior to the initiation of a monoclonal experiment. It is important that a screening strategy be defined beforehand which will result in the efficient detection of interesting hybridomas. Antibody-producing hybridomas need to be tested in the assay for which they will be used (i.e. if antibodies are to be used in Western blots, they need to be screened by Western blots) as soon as reasonably possible in the screening strategy. Services provided by the core on a typical project are immunization of animals, fusion, ELISA screening and two rounds of subcloning. The average cost for UNC investigators for the development of up to 8 hybridoma cell lines is approximately $3500-$5000 depending primarily on the number of clones selected for cloning and the number of ELISA plates used to screen the clones. *The standard immunization strategy requires ~150 µg/mouse of antigen, although there are alternative strategies which use less material. Please contact Bradley Bone to discuss these specific strategies. Monoclonal Antibody Services |
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