Tap into the extensive hybridoma development experience of Covance. A standard schedule rat hybridoma development project requires 2,950 µg of immunization antigen or 590 µg/rat for a total of 500 µg of ELISA screened antigen.
Rat hybridoma development production phases:
- Immunization (phase I)
- 5 Sprague-Dawley rats immunized
- immunogen emulsified in Freund's adjuvant at 3 week intervals (3 immunizations)
- ELISA titers of serum samples
- Rats hyper-immunized with low dosage boosts; used for fusion
- Fusion (phase II)
- Spleenocytes prepared from rat
- Fusion with hybridoma partner
- YB2/0 myelomas
- screen all potential clones by ELISA
- Top 30 clones expanded
- testing method of choice
- Cryopreserve for cloning
- Cloning (phase III)
- select clones
- identify best antibody producer clone with ELISA
- confirm media samples from top 10 antibody producing clones
- cryopreserve 10 clones in duplicate vials for temporary storage
- selection of subclone to be expanded; long term storage
- Stabilization (phase IV: optional, but recommended)
- Subclone selected clone; verify continued antibody production
- ELISA screen of subclones; media supplied
- Selection of clone/cryopreservation for long term storage
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