Monoclonal Antibody and Hybridoma Development Program
The identification of genetic biomarkers and pathways has revolutionized the process of drug discovery and created a flurry of novel therapeutic targets and methods. In today’s regulatory environment, it is critical to have high quality assay reagents to provide proof of concept, and more importantly, the ability to track the effectiveness of a therapy through clinical trials and beyond. MBS has partnered with top pharmaceutical companies, using 19 years of antibody development experience and proprietary protocols, to provide the best possible antibodies to monitor their therapeutic applications.
Typical Price Range For Hybridoma Development $7,300-$20,000
As a response to customer feedback, MBS offers a rapid immunization protocol resulting in a fusion at day 28 and your monoclonal cell line in 16-20 weeks. The protocol has been used with great success for over 4 years. A more traditional protocol, taking approximately 26 weeks for immunizations, is also available by request. Our hybridoma development team can help you to decide which method best fits your project parameters. The success of your project depends heavily on frequent communication with our staff throughout the project. Our goal is to help you get your diagnostic test or therapeutic assay to the market sooner.
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 Hybridoma 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
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. At this point, MBS is able to offer small amounts of purified antibody from the fusion products through a process called MultiPure. Learn more about MultiPure
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.
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.
Contact us today to learn more about our Monoclonal Antibody and Hybridoma Development Program.