QED Bioscience has over 23 years of experience in the development of hybridoma cells lines. We have made antibodies to large and small peptides, proteins, haptens, carbohydrates, bacterial and viral antigens, and plasmid DNA-encoded antigens. We understand and appreciate that every antigen is unique, so our hybridoma development programs are never "cookbook". We will tailor a program to your needs after we discuss important features of your antigens and the antibodies you require.
Features and Benefits:
- QED's programs are tailored to each client's specific requirements.
- Samples of antibodies are available to the client throughout the program for evaluation in their laboratory.
- All hybridomas and antibodies produced in our programs are the property of the client.
- Programs are set up in phases. Billing is at the beginning of each phase. If the program does not proceed to the subsequent phase, you are not billed.
- We require client authorization before proceeding from one phase to the next.
- Client receives regular progress reports as the program proceeds.
PHASE I: IMMUNIZATION
- Immunization of 5 mice. Additional mice available for an added charge.
- Protocol favors high affinity IgG responses.
- Titers of serum antibody to immunogen are monitored throughout the protocol; immune sera are available to the client for their evaluation.
- Testing of immune sera against potential cross-reactants is available for an added charge.
- Animals with highest serum-antibody titers are selected for fusion.
Estimated completion time: 10-14 weeks
PHASE II: FUSION
- Spleen cells from selected mice are fused with two different myeloma cell lines
- Fusion cultures are distributed to 6-96 well plates. Additional plates available for an added charge.
- Any remaining "fusion cocktail" is frozen per an in-house protocol for future use if needed.
- All antibody-positive parental cultures are expanded and frozen.
- Ten parental cultures are subcloned to ensure monoclonality. Additional parental cultures can be subcloned for an added charge.
- All antibody-positive subclones are expanded and frozen.
- Client receives all frozen cultures or can select which cultures they want to receive.
Estimated completion time: 2-3 weeks
PHASE III: ANALYSIS
- All parental fusion cultures are tested for antibody to immunogen, and all antibody-positive cultures are expanded.
- Expanded antibody-positive parental cultures are re-tested for antibody to immunogen. Testing against potential cross-reactants is available for an added charge. Culture supernatants are available to client for their evaluation.
- All subcloned cultures are tested for antibody to immunogen, and all antibody-positive subclones are expanded.
- Expanded antibody-positive subclones are re-tested for antibody to immunogen. Testing against potential cross-reactants is available for an added charge. Culture supernatants are available to client for their evaluation.
- Antibody isotypes are determined.
Estimated completion time: 6-8 weeks
PHASE IV: ANTIBODY PRODUCTION
- Antibody is produced either in ascites (5 mice per clone) or in vitro (150ml per clone) for 5 clones. Production can be performed for additional clones for an added charge.
- Client receives ascites or culture supernatant.
- Antibody purification available.
Estimated completion time: 3-4 weeks
Total Estimated completion time: 5-6 Months
Additional Hybridoma Services
Proven in-house immunization protocols and specialized assays are used to develop "internal image" anti-idiotype antibodies. Clients may provide their own monoclonal or polyclonal antibody to be used as the immunizing idiotype (Id), or QED will develop a monoclonal Id. When appropriate anti-Id responses are detected in immunized mice, selected mice are used for hybridoma production. Hybridomas are evaluated for production of anti-Id antibody, and additional assays to confirm internal imagery can be performed. Please contact a QED representative for a customized price quote.
Bifunctional Antibody Development
QED develops bifunctional monoclonal antibodies using our own proven patented technique. Different groups of mice are each immunized with different antigens. Sera from immunized mice are monitored for desired antibody responses. When appropriate antibody responses are attained, spleen cells from selected mice are combined and fused to myeloma cell lines, and resulting hybridomas are subjected to dual analysis for reactivity with each antigen. Ultimately, functionality is confirmed by successive affinity separations. Costs of Immunization Phase (Phase I) and Analysis Phase (Phase III) are approximately twice the costs described above; costs of Fusion and Antibody Production are the same.
Is Something Missing?
We have described the services most frequently requested from QED. If we did not include an antibody-related service that you need, call us - chances are we can help. If you need any other information, or if you would like to order any of our services, we will be happy to assist you.