Monoclonal Antibody Production
Services request form for monoclonal antibody production
All the procedures-protocols used and the care of experimentation animals is performed according to the specifications established in Real Decreto 21/10/2005, on "Protection of animals used for experimentation and other scientific purposes”.
The antigen will be provided by the customer. The amount of antigen required varies depending on the nature of the antigen and will be agreed previously with the client. We recommend the following amounts:
- Recombinant proteins: around 1.5-2.0 mg.
- Synthetic peptides: around 3.0 mg.
Standard Protocol for the Generation of Monoclonal Antibodies
The standard protocol for the development and production of monoclonal antibodies will last a period of 4-6 months. It includes the following modules:
- Preliminary meetings to discuss the project strategy.
- Immunization of 5 BALB/c mice
- Screening and selection for positive clones by ELISA, or other immunoassays, followed by two cloning steps by end-point dilution.
- Customer delivery of around 1 ml of culture supernatant from selected positive clones for functional assessment of monoclonal antibodies under the customer conditions.
- Growth / Expansion / Cryopreservation of selected hybridomas
- Further hybridoma characterization: IgG subclass determination (Isotyping assay) and testing for mycoplasma.
The customer will receive:
- Mouse pre-immune serum, around 500 μl.
- For each selected hybridoma (up to three): three cryovials and 15ml of the culture supernatant will be provided.
- Information about the antibodies.
Intellectual Property: The resulting hybridomas will be owned by the customer.
- Protein G purification
- Isotyping assay
- Biotin or Peroxidase coupling
- Liquid N2 storage (1 year)
- Hybridoma cell culture
- Functional validation in ELISA, Western Blot, IHC, IF and/or flow cytometry techniques