Bio-Synthesis's success rate in Custom Hybridoma Development for
Monoclonal Antibodies from fused cell line is greater than 95%. Relying
on our proprietary Antigen Identifier design tool, flexible peptide synthesis platform
and proprietary SP-Max™ adjuvant, we deliver high quality monoclonal antibodies
in a short turn-around time at affordable cost.
Each of our monoclonal antibody development packages include:
The following quality control measurements are employed to create the highest quality
monoclonal antibodies commercially available:
There has been much discussion about using recombinant or fusion proteins versus peptides as antigens. IgG antibodies work for a wide variety of applications, such as western blot, immunostaining, immunoprecipitation, and more. When obtaining IgG antibodies, a protein antigen works better for custom monoclonal antibody development. The immune system of a mouse responds better to proteins due to the higher number of linear and conformational epitopes. Ultimately, the more diverse of an immune system response usually results in antibodies against multiple locations on the protein. Each of these protein locations has unique properties.
Peptide antigens are usually only linear in nature and also have fewer epitopes. To increase the total number of epitopes available for the mouse immune system to respond to, it is recommended to use peptides of 20-30 residues in length. Increasing the immune response will increase the likelihood of success of your custom monoclonal antibody project. Going with shorter peptide sequences of less than 20 residues in length usually only works for ELISA.
When submitting a protein antigen for monoclonal development, it is highly recommended to cleave all GST tags or other large fusion partners from the protein. Usually, large fusion partners are highly immunogenic. If they are not removed before immunization, a dual ELISA screening process must be performed to ensure positive responses are results of protein and not to the fusion partner. For this process, high ELISA titers are screened against the protein, while low ELISA titers are screened against the fusion partner.
Bio-Synthesis offers a variety of Hybridoma Development Monoclonal Antibody Production
Services. Customer personalized packages are also available. To initiate
a custom contract monoclonal antibody service project, please contact our Technical
Service Department at: (800) 227.0627 or contact us online. A project manager will
be assigned to help you select or design the best service for your specific needs.
Available service for pre-immunization antigen processing and modification
Standard Protein (mAb1000) or Standard Peptide mAb (mAb3000) packages : Bio-Synthesis, Inc. will guarantee up to three (3) ELISA positive hybridomas to the immunogen with 1 x 105 cells of each clone to either a protein antigen or a peptide antigen BSI selected and synthesized. Because of the nature of hybridomas, and antibodies in general, it is not possible to guarantee that the clones will work for applications other than ELISA, or that a clone will work for multiple applications.
Premium Protein mAb packages (mAb2000): Bio-Synthesis, Inc. will guarantee up to three (3) positive hybridomas for the screening application (western blot or immunocytochemistry) selected by the client with 1 x 105 cells of each clone.
Premium Peptide mAb packages (mAb4000): Bio-Synthesis, Inc. will guarantee up to three (3) ELISA positive hybridomas to the peptide antigen with 1 x 105 cells of each clone. Bio-Synthesis cannot guarantee reactivity for a specific application with peptide antigens; if the first fusion does not yield clones that are positive for the screening application chosen by the client (western blot or immunocytochemistry), a second fusion will be performed at no charge. Bio-Synthesis can terminate the project if the second fusion is not successful in yielding clones for the selected screening application and will not invoice phase III or will refund phase III if paid and client will only pay for peptide synthesis, conjugation, immunization, and one fusion.
Researcher Screening of Positive Wells: In order to improve the chances of obtaining antibodies suitable to
your needs, clients have the option of screening the cultured supernatants from up to 100 positive wells
following fusion. If this option is ordered, client results must be forwarded to Bio-Synthesis within 5 days of shipping of the supernatant samples.
After this time a non-refundable per diem maintenance fee will be charged to
your account. Because of the time critical nature of cloning, selections received after the 5day period will
void any expressed or implied guarantees. Please note Bio-Synthesis cannot guarantee the continued reactivity
seen in supernatants throughout sub-cloning.
Antibodies that work well in one application may not work in another. We can, however, guarantee that each clone will recognize the immunogen in ELISA as this is our primary method of screening. The success of a project is dependent upon the immunogen, screening methods for hybridomas, etc. We highly recommend screening positive wells, especially if a peptide immunogen is used as this will increase the chances of developing a cell line that produces monoclonal antibodies that work in your target application. Additional positive clones may be available at additional cost.
We cannot guarantee the success of a project where the customer provides the antigen; however it is our experience that monoclonal projects using a recombinant protein yield monoclonal antibodies that have a higher recognition rate to the native protein than by using a peptide immunogen. The customer must also be aware that a customer-provided immunogen may not yield the immune response required for fusion. If the customer provides the immunogen there will not be a refund for phase one if there is insufficient titer for fusion following the 6th test-bleed in the immunization protocol.
If there is a successful fusion with no resulting ELISA-positive clones, phase one will not be refunded, although we will perform a second fusion at no charge. If the customer is not satisfied with the positive wells at screening or final clones they may opt to pay for another round of fusion and/or sub-cloning at additional cost.