ImmunoPrecise’s proprietary Rapid Prime Method in mice identifies positive monoclonal antibodies in as little as 32 days. While this method also uses Single-Step cloning to accelerate the process, it is especially good for generating anti-idiotypic antibodies and has also been extremely useful for generating monoclonal antibodies against conformational epitopes. Other sources of immunogens include conserved or smaller proteins, peptides, and whole cell bacteria.

Project Phases Timeframe
I. Immunization

  • Immunization of 4 female BALB/c mice
  • Antigen requirements:
    • Please refer to FAQ for more details
    • Antigens in solution must be in PBS, neutral pH, at a concentration of 0.5-2mg/mL and contain nothing harmful to animals (no preservatives)
19 Days

Rapid Prime Phase 1

II. Fusion and Selection of Hybridomas

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

Rapid Prime phase 2

III. Screening 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

Rapid Prime 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

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.