Custom hybridoma and antibody production

Custom hybridoma and antibody production options for rat and mouse monoclonal antibodies

We offer options for the development of both rat and mouse hybridomas for the production of custom monoclonal antibodies from synthesized peptide or recombinant protein antigens.

  • Design your own project—Select one of our robust protocols for immunizing mice or rats, the desired number of fusions, add subcloning, and select one of the production options.
  • Get advice from expertsContact our experienced scientists and get their professional advice on how to design a project best suited to your specific needs.
  • Defer costs—The antibody development schedule is divided into five phases and payment stages, enabling aspects of the project scope to be changed in-process.
  • Use our full-service integration—Use our additional services, such as antigen development, to accomplish a phase of development when you do not have the resources to perform it yourself.
Details and additional information

An entire project for monoclonal antibody development, production, and purification requires approximately 6 months to complete. The process is divided into 5 phases (milestones), which are invoiced after the completion of each phase. These milestones represent exit points for discontinuing the project if results do not meet your specific expectations.

Antibody labeling can be added as a sixth phase to the antibody development project. Our standard methodology is as follows:

antibody development project
Phase 1: Antigen preparation

Customers may provide highly purified (>90%) protein or peptide antigens that are ready for injection. Alternatively, we provide services for protein production, peptide design, peptide synthesis, and peptide conjugation to carrier protein for use as the immunogen.

Phase 2: Immunization and ELISA titration

Animals are immunized and test-bled over a 6-week period (one primary injection and two booster injections). A titration ELISA is performed with each test-bleed. ELISA results and crude antisera are sent to the customer for evaluation. See mouse protocols or rat protocols for additional details.

Phase 3: Fusion and screening for positive supernatants

The best-responding animals are boosted twice more; then their spleen cells are harvested for hybridoma fusion (10 plates per fusion). An ELISA assay is used to screen the positive supernatants, 1 to 2 mL of which are sent to the customer for evaluation. Cell lines are preserved (expanded and frozen) and stored during customer evaluation. See monoclonal antibody ELISA testing for additional details.

Phase 4: Subcloning before production

Parental cell lines can be subcloned through limiting-dilution cloning to obtain daughter cell lines. Subcloning is important for the long-term stability of clones and to ensure that cells are truly monoclonal and maintain clonality over their production life. Multi-cycle complete clonality cloning involves performing multiple rounds of limiting-dilution subcloning until all wells are growing at the same rate, are secreting antibody at the same concentration, and have the same confirmed isotype.

Phase 5: Production and purification

Monoclonal antibodies are produced in cell culture in roller bottles to yield antibody-containing supernatants. Each liter of cell culture yields approximately 10 to 30 mg of antibody. Low-endotoxin and serum-free culturing are available on request, as well as high-volume production.

Depending on the isotype of the monoclonal antibody, affinity purification is performed using Protein A or Protein G resin.

Phase 6: Labeling

We offer a wide variety of custom fluorescent antibody and protein labeling options.