ImmunoPrecise’s Standard Method monoclonal antibody development service in Mice and Rats is fast and efficient. IPA’s single-step cloning method shaves weeks off of the traditional time line for hybridoma production. IPA custom develops monoclonal antibodies to an array of antigens including peptides, modified peptides, recombinant proteins, DNA and small molecules.

Project Phases Timeframe
I. Immunization

  • Immunization of 4 female BALB/c mice or 2 female F344 (SAS FISCH) rats
  • Test bleed is taken 7-10 days after 2nd boost
  • Antibody titre is evaluated by ELISA on screening antigen
  • Best responding 2 mice or best responding rat is chosen for fusion
  • Final boost (IV or IP) is given 4 days before fusing
  • Antigen Requirements:
    • Please refer to FAQ for more details
    • Antigens in solution must be in PBS, neutral pH, at a concentration of 0.5-2.0mg/mL and contain nothing harmful to animals (no preservatives)
63 Days

Standard Method Phase 1

II. Fusion and HAT Selection of Hybridomas

  • Lymphocytes from 2 mice are pooled, purified then fused with SP2/0 myeloma or
  • Lymphocytes from 1 rat are purified then fused with X63Ag8 mouse myeloma
  • Fused cells are grown on HAT selective Single-step cloning media for 10-12 days
  • Up to 948 individual hybridoma clones are transferred to 96-well plates and grown up for 4 days
16 Days

Standard Method phase 2

 III. Screening and Expansion of Hybridoma Clones

  • Primary screening of hybridoma tissue culture supernatants by indirect ELISA using screening antigen
  • Positive clones are further tested by ELISA on screening antigen and negative control antigen (human transferrin) to confirm Ig secretion, specificity and to rule out false positives
  • Positive clones are isotyped and subcloned to identify stable expressing clones (“Stability Period”)
  • Tissue culture supernatants can be shipped to the client for testing during the “Stability Period”
32 Days

Standard Method phase 3

 IV. Subcloning and Cryopreservation

  • Client can choose up to 5 positive clones with the option to purchase more
  • 5 cryovials of each chosen clone plus 40mL of overgrown cell culture supernatant will be shipped to the client by courier on dry ice upon completion
  • 2 cryovials per clone will be stored in liquid nitrogen at IPA for a period of 1 month
  • Option to store cryovials long term at IPA is available, please click here for more information
35 Days

Standard Method Phase 4

V. Antibody Concentration and Purification (Optional)

  • IPA offers in vitro and in vivo techniques, please refer here for more information
  • IPA offers a variety of purification techniques, please refer here for more information
4-7 weeks

For a quote please call us at 1-250-483-0308, or via email at contact.ipa@immunoprecise.com. You may also reach us by completing our online form.