The Hybridoma core lab provides services and products related to the production of monoclonal antibodies. Basic procedures include mouse immunization with immunogen provided by the requestor. Subsequent boosts and bleeds are done to check antibody titer in the animal. Spleen cells are fused with a non-immunoglobulin secreting mouse myeloma cell line. Hybridomas are cultured in standard selection medium and are screened by a variety of methods by the requesting lab, or by the Hybridoma core by ELISA. Primary cultures of hybridomas are subcloned by cell sorting and screened again. Once a stable hybridoma cell line is produced and its isotype and titer are determined, the core lab can produce ascites in vivo or use in vitro methods of antibody production. Cells are frozen at each stage of production and are kept in two locations.

In addition, the Hybridoma core maintains an inventory of established hybridoma cell lines, as well as primary cultures of endothelial cells. For more information about inventoried cell lines and availability, contact the core staff at ihernandez@versiti.org.

To access Hybridoma services or monoclonal antibody production from existing hybridomas, please request a consultation with the core staff via the iLabs platform under the “Request Services” and “Hybridoma” tabs. A detailed list of specific services is available under the “Request Services,” “Services” and “Hybridoma” tabs. Services include:

• Immunization of mice (Balb/c or Nude)
• Test bleeding and screening of immunized mice by ELISA
• Splenocyte-myeloma fusion
• Screening of primary hybridoma cultures by ELISA
• Subcloning of primary hybridoma cultures by cell sorting
• Screening of subclones by ELISA
• Antibody titer and isotype measurement
• Expansion and cryopreservation of clonal hybridomas
• Ascites production in Balb/c mice
• In vitro antibody production in bioreactors

For immunization, the user will provide approximately 1 mg of purified protein or peptide coupled to a carrier protein. Generally, fusions are done with Balb/c mice. The Hybridoma core lab offers related services as time and demand permit. Ascites antibody production from in-house hybridoma or ATCC cell lines may take three weeks to three months, depending on demand. If there is a sufficient supply, 3-15 ml of ascites can be provided from freezer inventory within 24 hours. 

New fusions are on a first-come, first-served basis. Preference is given to Versiti Blood Research Institute investigators. Generally, the requesting lab needs to provide about 1 mg of purified protein or peptide coupled to a carrier protein. The immunization period is typically 3-4 months. Screening, cloning, subcloning hybridoma and freezing of cells can take an additional 3-6 months. All hybridoma screening methods are to be worked out prior to fusion date.