The Hybridoma Core Facility was established to provide investigators at the Lerner Research Institute with a resource for monoclonal antibody production. The facility also provides consultation services to assist the investigator in all aspects of monoclonal antibody production. The services are performed under strict QC guidelines appropriate for antibodies that may have commercial potential.
There are two phases of monoclonal antibody production: the development of the hybridoma cell line and production of the monoclonal antibody from the established hybridoma cell line. Currently, the Hybridoma Core offers two methods for monoclonal antibody production from hybridoma cell lines, tissue culture in T-Flasks and the Integra Flask system. Virtually all hybridoma cell lines will grow in vitro; the choice of method depends on the amount, concentration, and purity of antibody required.
Polyclonal antibody production is offered on a routine basis, using peptide or protein antigens.
One rabbit per antigen. Includes animal purchase and board, 5 injections, 2 production bleeds and one final bleed in a 90 day protocol. Time course can be extended for additional fee based on animal board and labor charges. The goal is to provide a total of 80-100mls of serum, but this is not guaranteed. Allow 1-2 weeks for animals to arrive.
We can purify monoclonal or polyclonal antibodies using a Protein G column or an epitope specific affinity column. The affinity column will be sent to the investigator and can be used for future purifications.
The Hybridoma Core uses a static cell culture system (the Integra Flask) to produce high concentration (>.5mg/ml) monoclonal antibodies. This can be done in serum-free or using ultra-low IgG/IgM serum. Yields can be as high as 100mg/month/flask. The Core has produced as much as 5 grams of antibody by this method.
The Hybridoma Core will inject 4 mice 3 times plus a final boost. Serum titer is measured by ELISA. The spleen from the best mouse is used for the fusion. Typically, 10-15 plates(96 well) are used in the fusion. Positive hits are detected by ELISA in the Core - or - the supes can be sent to the investigator for screening. Up to 100 hits can be selected for a 2nd round screening assay - by ELISA in the Core or by the investigator in a more specific functional assay. 1-10 highly specific hits can be cloned.
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Hybridoma Core has not received any endorsements.