The Monoclonal Core was established in 2000 and has been directed by Dr. Daniel Cawley since 2004. The VGTI Monoclonal Core offers two basic services: the generation of new monoclonal antibodies and the scaleup production and purification of antibody from existing cell lines (a stand -alone phase IV). A project is typically done in four stages.
Stage One Immunization
4 mice are given 2-3 injections with protein antigen. Antisera titers are determined to identify animals for fusion and to set up an ELISA screening assay.
Stage Two Fusion and hybridoma screening
Spleen cells from 2 mice are fused with mouse myeloma cells to generate hybridomas. After ELISA screening, cell supernates are delivered to the client for testing.
Stage Three Cloning and characterization of hybridomas
Parent hybridomas are limit dilution cloned, providing for stable growth and antibody production and monoclonality. Several vials of frozen cells are made available to the client.
Stage Four Antibody scaleup production and purification
In vitro scaleup in bioreactors provides antibody in the 50 mg range. Antibodies are purified to greater than 90% using two chromatography steps. A list of existing cell lines for Ab production is available upon request.
FEEs
|
