Monoclonal Antibody and Hybridoma Development Program
MBS is proud to offer two effective schedules for Hybridoma Development. The first, the standard immunization protocol, takes approximately 26 weeks. The second, increasingly popular method, is our Rapid Immunization Protocol which is based on a 28 day immunization schedule and produces a hybridoma cell line in approximately 16-20 weeks. Our experienced sales and scientific staff can help determine which method best fits your needs and project parameters. The success of both protocols depends heavily on frequent communication throughout the project.
The Hybridoma Guarantee explains our commitment to quality.
Phase 1 Project Initiation
This phase provides you with the opportunity to discuss the overall goals of the project and tailor our protocols to meet your requirements. To begin this stage, please fill out a Monoclonal Antibody Development Checklist. This form will supply MBS with all of the non-proprietary information and help us to get started with your development.
Hybridoma initiation checklist
Phase 2 : Immunization of your Mice or Rats
An antibody response is generated in mice or rats to your specific immunogen. Up to 8 animals can be immunized if required. A standard project requires 3-4 mgs of immunogen, however if less is available the rapid hybridoma development can be modified. Immunogens less than 20kD should be conjugated. We recommend conjugating 2mgs to KLH for immunizations and 2mgs to BSA for screening of test bleeds and supernatants. After 28 days, the selected animal goes to fusion.
Important Immunogen Factors To Consider
Pricing for Immunization Phase: $2,400-$3,500
Phase 3: Fusion
Spleen cells from the selected animal are fused with myeloma cell line to develop a hybridoma that secretes a specific antibody. Once your mice or rats are ready for fusion per ELISA screening results, a fusion is performed. The fusion is plated and screened. The primary screening is a direct ELISA against your antigen to capture all positive fusion products. The positive fusion products are scaled up and screened again. The secondary screening differentiates between IgG and IgM clones, and verifies that the selected positive fusion products are still producing antibody prior to cryopreservation. You will choose the number of fusion wells for scale-up. Not all wells that are scaled up will survive or continue to produce specific antibody. The supernatants of the positives are shipped to you for further testing. If you require small amounts of purified antibody for testing 20_g of each positive can be provided by our Block Purification Process.
Pricing for Fusion Phase $8,000 - $11,000
Phase 4: Subcloning
A monoclonal antibody-producing cell line is isolated from selected parental clone. MBS performs subcloning to render a hybridoma fusion product, or cell line, monoclonal. The selected fusion clones are subcloned by limiting dilution. MBS has three proprietary approaches to subcloning, which are used depending on the stability of the clone. Once colonies are established, the wells are screened by a direct ELISA assay. Clones from each parental are selected for freeze down and storage. The supernatants are shipped to you for further evaluation. This phase takes 3-4 weeks.
Pricing for Subcloning Phase $2,000 - $5,000
Phase 5: Antibody Production
Sufficient antibody is generated through ascites or in vitro production to allow you to assay in your specific application.
For cell lines developed in Balb/c mice, we typically use 10 Balb/c mice to produce Ascites. If your cell line was developed in rats, we will produce a Spinner Flask production or run a nude mice ascites production. The ascites or spinner flask production can be purified by Protein A or Protein G when the antibody is an IgG. The final monoclonal antibody is shipped to you within 7-8 weeks of selecting a subclone.
Pricing for Antibody Production, Pilot Scale $800 - $1,500
Contact us today to learn more about our Monoclonal Antibody and Hybridoma Development Program.