HYBRIDOMA CORE FACILITY
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Director:
Clemencia Colmenares, PhD
Staff, Cancer Biology
colmenc@ccf.org
Link to the Hybridoma Core Facility home page here.
Services
The goal of the Hybridoma Core is to provide a cost-effective resource for generating and producing monoclonal antibodies to novel antigens that will enable investigators to discriminate among the proteins relevant to their research. Highly specific monoclonal antibodies are an essential resource in identifying the levels of expression and specific modifications of proteins involved in all the functions that control cell growth, cell differentiation, and cell death, and therefore constitute a critical tool in all aspects of cancer research that involve specific proteins.
This Core provides both routine services (such as the expansion of hybridomas to produce concentrated, serum-free monoclonals) and also customizable services (such as the generation of hybridomas). For the larger projects, the core works with the investigator to customize, for example, the selection of mouse strains to be immunized in order to improve the likelihood of developing antibody titers; or, in the interest of economy, the core also trains researchers if they wish to take over the project and perform the subcloning themselves.
The services provided by the Hybridoma Core include:
- The generation of mouse monoclonal antibodies, as a complete project or broken down into 3 separate steps: injection of the antigen, fusion and selection, and cloning and screening
- Production of high-concentration antibodies from hybridoma cell lines cultured in serum-free Integra flask cultures
- Generation of polyclonal antibodies in rabbits
- Purification of polyclonal antibodies on antigen affinity columns (new service)
- Production of monoclonals from existing cell lines, as well as cryopreservation, and storage of lines
- Training of researchers who wish to perform parts of the project such as the selection, subcloning, and screening of potential hybridomas
Fees
| Prefusion | The investigator will provide the antigen and the HCF will prepare it for injection. Immunization and serum collection from mice for screening by investigator. Includes animals, labor and supplies. This phase goes at least 8 weeks or until desired titer is reached. | $568 |
| Fusion | Fusion of spleen cells from chosen mouse to myeloma cell line. Plating of fused cells and collection of media samples for screening by the investigator. Includes labor and supplies. This phase goes 4-6 weeks. | $1703 |
| Cloning | Limiting dilution cloning of five chosen hybridoma cell lines. Collection of media samples for screening by the investigator. Minimal scale up and preparation of frozen stocks (5 vials) of clones are completed by HCF. Includes labor and supplies. This phase goes 4-8 weeks. | $1703 |
| MAb production of mAb from cell lines by cell culture methods | Cell culture expansion of cloned hybridoma cell lines and collection of conditioned media. Additional concentration and purification of the mAb to be done by investigator. Mycoplasma testing is recommended for cell lines not made by the facility. | $55/100mls |
| Freezing Cells | Cells from culture expansion associated with mAb production can be frozen down for storage in Liq N2. The freezing media is 90% FBS:10% DMSO and the cells are at a concentration of 2-4 x 106 cells/ml. **If a vial is brought only for expansion and freezing, there is an additional $25.00 tissue culture charge. | $34/5 vials |
| Integra Culture System for production of mAb from cell lines | This system is intended for production of 30-60mg of mAb per month in the smaller system and 100-200mg of mAb in the larger system. Average concentration is 1.5mg/ml. The production schedule average is 8 weeks. | CL 350: $454 CL 1000: $908 |
| Polyclonal Antibody Production | This service will be offered on a limited basis depending on available housing. One rabbit per antigen. Includes animal purchase and board, 6 injections, 1 test bleed, 3 production collections, final bleed and labor for 120 days. Time course can be extended for additional fee based on animal board and labor charges. The goal is to provide a total of 100mls of serum, but this is not guaranteed. Allow 1-2 weeks for animals to arrive. | $551 (short protocol, 1 rabbit) $1212 (long protocol, 2 rabbits) $673 (long protocol, 1 rabbit |
| Antigen Conjugation | Peptide conjugation to KLH by a glutaraldehyde protocol. | $171 |
| Liquid N2 storage | Storage for cloned cell lines. Price is for each cell line. The stored cell lines must be mycoplasma free. | $16.50/mon |
Contact for Information
| Location: | Lerner Research Institute, NB1-25 |
| Director: | Clemencia Colmenares, PhD |
| Phone: | 216/445-6613 |
| E-mail: | colmenc@ccf.org |
| Manager: | Earl Poptic |
| Phone: | 216/445-6635 |
| E-mail: | poptice@ccf.org |