Antibody Engineering and Technology Core resource description
The Antibody Engineering and Technology (ABET) core facility, established in 1983 as the Lymphocyte Culture Center, provides all investigators at the University of Virginia access to advanced technology, and cost effective monoclonal antibody isolation and engineering services. The core gives researchers access to the most current technology and expertise in all aspects of antibody engineering techniques including construction and selection of lymphocyte-myeloma hybridomas for the production of monoclonal antibodies. The core provides advanced techniques for shaping antibody molecules including; (1) Antibody hybridoma sequencing from mouse and rabbit species; (2) construction of small antibody fragments (scFv, Fab); (3) chimeric and humanized antibody and affinity maturation; (4) modification of multivalent antibody format (bispecific); (5) production of antibody using hybridoma cell, CHO, HEK293 cells and E.coli expression and purification using affinity chromatograpy. The core can also provide 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 specific monoclonal antibodies. The core has also provided research support services to other academic institutions and companies, and to federal agencies such as the NIH, the FBI, and USAMRIID.
The center occupies a modern laboratory of approximately 1270 square feet equipped with cell culture incubators, laminar flow hoods, centrifuges, inverted and standard microscopes, automated microplate washer and ELISA plate readers, automated HPLC system, and PCR. We also have facilities for the cryopreservation and long-term storage of established hybridomas and myeloma cell lines in liquid nitrogen cell banks. The core is currently staffed by a director and two laboratory specialists advanced.
The core will immunize animals, collect sera and develop ELISA assay strategies appropriate for screening large numbers of hybridoma culture supernatants and animal sera. Once an appropriate assay system has been developed (either in the core or in the individual researcher’s laboratory) to demonstrate the successful immunization of the necessary animals, fusions are scheduled on a first come first serve basis. The core 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 including bulk expansion of clones, isotyping and sub-isotyping of monoclonal antibodies, bulk monoclonal antibody production in vitro using single use bioreactors (service does not include IgG3 and IgM serotypes), and monoclonal and polyclonal antibody purification by affinity chromatography on recombinant Protein G columns. The core 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.