MONOCLONAL ANTIBODY DEVELOPMENT
Each of our monoclonal antibody development packages include:
- Immunization of antigen into 5 BALB/c mice
- Screening of test bleeds against immunizing antigen to isolate a mouse with the high IgG titer
- Fusion of one mouse's spleen to the FO myeloma cell line
- Screening of positive wells following fusion for IgG specific clones using immunizing antigen
- Two sub-cloning steps to ensure a completely monoclonal colony
- Cell line stabilization of up to 3 cell lines
- Isotype report on each final clone provided, confirming Ig class
- Cell line banking of backup vials for 1 year following final clone shipment
A summary of our packages can be found in our Monoclonal Antibody Development Information Sheet (PDF).
Additional options and services include:
- Peptide synthesis (with antigen design assistance)
- Multiple ELISA screening (i.e.: screen positive hybridomas against GST to avoid selection of clones secreting antibodies against GST and not the protein of interest)
- Researcher screening of positive wells following fusion; up to 7-day screening time
- Additional Western blot or immunocytochemistry screening
- Purchase of additional clones
- Ascites production and purification
- In vitro production and purification
- Cell line isotyping
- Cell line banking
- Cell bank generation
Ready to order? Just fill out the following Monoclonal Antibody Development Order Form (PDF).
Protein vs. Peptide Antigens:
We are frequently asked about the merits of using recombinant or fusion proteins versus peptides as antigens. To obtain IgG antibodies that work for multiple applications (i.e.: western blot, immunostaining, immunoprecipitation, etc.) a protein antigen works much better for custom monoclonal antibody development. Proteins have a much higher number of linear and conformational epitopes for the mice's immune systems to respond to. The more diverse immune response generally results in antibodies against many different locations on the protein, each of which has unique properties. Peptide antigens have fewer epitopes and are normally only linear in nature. Using longer peptides (~20-30 residues in length) increases the total number of epitopes available for the mouse immune system to respond to and increases likelihood of success of your custom monoclonal antibody development project. Our peptide-antigen packages include 25 residues, to help increase the project's success. Shorter peptide sequences (fewer than 20 residues in length) often work only for ELISA.
Fusion Partners (protein antigens):
If you are submitting a protein antigen for custom monoclonal antibody development, we highly recommend cleaving large fusion partners such as GST tags. These tags tend to be highly immunogenic, and if they are not removed prior to immunization, a dual screening process must be performed. This ensures positive responses are to the protein, not the fusion partner. To do this we perform dual ELISA screening, screening for high ELISA titers against the protein and low ELISA titers against the fusion partner.
Custom Monoclonal Antibody Development Antigen Requirements:
- Purity: Antigens should be as pure as possible (>90% is recommended). An antibody can be made against impurities just as easily as against the protein itself.
- Solubility: Antigens need to be soluble for screening.
- Antigen Types: ProSci Incorporated can work with protein, enzymes, and peptides. At this time we do not accept cells, viruses, or bacteria as antigens for custom monoclonal antibody production.
- Antigen Quantities: From start to finish ProSci needs ~5mg of antigen. 1mg is used for immunizations and 4mgs are used for screening. We can start a project with as little as 0.5mg, and additional quantities can be supplied throughout the project
- Conjugation of Antigen: Small antigens (such as peptides or antigens with a molecular weight of 10kDa or less) need to be conjugated to a carrier protein to have a large enough size to initiate a strong immune response. ProSci requires 2-3mg of material to conjugate to the carrier protein (or 1mg conjugate if provided by researcher), and an additional 4mgs of antigen for screening
Monoclonal Antibody Development Terms and Conditions (PDF).
If you have any questions regarding our monoclonal antibody development service please contact us at abservices@prosci-inc.com, or by calling toll-free 888-513-9525.