Hybridoma Shared Research Facility

The Hybridoma Center has extensive experience in the development, production, purification and processing of monoclonal antibodies. The facility is conveniently located on the 16th floor of the Annenberg Building of the Mount Sinai School of Medicine. The animals that are used for the production of hybridomas are maintained in a site accredited by the American Association for Accreditation of Laboratory Animal Care (AAALAC).

A custom program proposal will be generated for each client to maximize the probability of obtaining the desired monoclonal antibodies while minimizing dollar risks to the client.

Procedures
The Hybridoma Center provides a complete hybridoma development program designed to produce hybridoma cell lines that secrete monoclonal antibodies specific to your antigen.

Enough antigen must be provided for the immunization of animals and for screening of the fusion. The antigens can be fusion or other proteins, protein linked peptides or other immunogenic products.

Immunization: Generally, three BALB/c mice receive primary immunizations in the foot pads with the antigen combined with Freund's Complete Adjuvant. The animals get a secondary immunization intraperitoneally (IP) approximately three weeks later with the antigen combined with Incomplete Freund's Adjuvant. Ten days later, mouse serum will then be screened via radioimmuno assay (RIA) or enzyme linked immunosorbant assay (ELISA) to determine the immune response.

If the serum titers are deemed acceptable by both the Hybridoma Center and the client, animals will then receive a final boost three to four days prior to a fusion intravenously (IV) via the tail. If the serum titers are determined to be unacceptable, other alternatives will be discussed.

Fusion: The fusion entails splenocyte fusion with a myeloma cell line. The Hybridoma Center uses as a fusion partner, SP2/0, which is a nonsecreting BALB/c myeloma.

BALB/c mice are used for immunization unless a different strain is requested.

Screening: Hybridoma cultures are screened to identify colonies secreting monoclonal antibodies specific to the target antigen. A negative antigen is encouraged to ensure that "false" positives are not selected.

Cloning: After consultation with the client, positive hybridomas, which from then on will be termed, mother hybridomas, will be cloned. After an approximate period of eight to ten days, the colonies will be screened against the antigen.

Isotyping of the positive hybridomas will be included.

Products and Services

Antibody Purification
Supernatant from a specific hybridoma will be grown up to approximately 500 ml. Then it will be affinity purified according to its specific isotype.

Hybridoma Supernatant

A volume of 500 ml of supernatant is the typical amount of hybridoma supernatant grown. However, if necessary, more can be produced.

Freezing and Storage of Hybridomas:

Hybridomas made by the Hybridoma Center can be frozen and stored.

Pricing

For specific pricing please contact us at 212-241-6401 or e-mail: erin.petersen@mssm.edu

Hybridoma Center Services

1. Hybridoma Production
   Basic package includes:
   • STEP ONE
     • primary immunization
     • secondary immunization
     • preliminary screening of mouse serum titers for immune response
     [If the Hybridoma Center in conjunction with the client decides to proceed to step two of hybridoma production, it should be noted that the package price includes this initial cost.]
   • STEP TWO:
     • fusion
     • screening for colonies secreting monoclonal antibodies specific to antigen
     • cloning
     • freezing of monoclonal antibodies
     • isotyping
   Other options are available. Specific steps of the package can be done individually. Please consult with the Hybridoma Center representative.
2. Hybridoma Supernatant
   500 ml of supernatant
3. Purification of Supernatant
   Based on 500 ml volume for supernatant.
4. Freezing of Hybridomas
   The freezing of 3 vials of hybridomas and one year of storage.

Questions and Comments

Please send e-mail to erin.petersen@mssm.edu