Rat Hybridoma Development

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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Immunology Services

Tel: 800-345-4114
Tel: 717-336-4921
Fax: 717-336-3481

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