Monoclonal Antibody Development

ProSci can help you develop custom monoclonal antibodies and we understand researchers have varying needs for levels of involvement, depending on the target. We have developed monoclonal antibody development packages for different levels of involvement in the project, from researcher screening (you screen positive wells following fusion for clones of interest) to ProSci sending western blot and immunocytochemistry positive final clones. We offer custom monoclonal antibody development packages for both protein and peptide antigens. Our screening options include:

• ProSci sends final clones, guaranteed ELISA positive. (no additional charge)
• Researcher screening of positive wells, following fusion. (starting at $500)
• Western Blot screening: ProSci screening of positives during Phase 2 (Fusion) and Phase 3 (Sub-cloning) for Western Blot positive clones.
  • Protein antigens: guarantee of up to 3 western blot positive clones. ($2200-$2600)
  • Peptide antigens: cannot guarantee reactivity, but >70% success rate; see Terms & Conditions for more details. ($2200-$2600)
• ICC screening: ProSci screening of positives during Phase 2 (Fusion) and Phase 3 (Sub-cloning) for immunocytochemistry positive clones
  • Protein antigens: guarantee of up to 3 immunocytochemistry positive clones. ($2600)
  • Peptide antigens: cannot guarantee reactivity, but >70% success rate; see Terms & Conditions for more details. ($2600)
• Western blot & immunocytochemistry screening: ProSci screening of positives during Phase 2 (Fusion) and Phase 3 (Sub-cloning for Western Blot AND immunocytochemistry positive clones. ($4200)

More pricing information can be found at the mouse monoclonal antibody developement page.

ProSci’s fusion success rate for custom monoclonal antibody developement is greater than 95%, using techniques developed in-house. All work is performed by skilled technicians on premises in our San Diego, California laboratories. Your custom monoclonal antibody production project, and can be scaled up to almost any magnitude. We can screen against multiple antigens (additional charges apply) to develop the specific clone you need. ProSci has repeatedly proven our abilities to produce hybridomas against difficult targets.

We use 5 Balb/c mice as a production strain for hybridoma development. Balb/c is a very common species used throughout the industry for monoclonal antibody development due to their large spleen to body size ratio. This yields a higher number of spleenocytes at fusion.

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 developement, 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. This additional service is available at $2400 per additional screening.

Custom Monoclonal Antibody Development Antigen Requirements:

• Purity: Antigens should be as pure as possible. 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, peptides, and other small molecules. 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 ~5 mg of antigen. 1mg is used for immunizations and 4mgs are used for screening. We can start a project with as little as 0.5 mg, and additional quantities can be supplied throughout the project
• Conjugation of Antigen: Small antigens (such as peptides or antigens with a molecular weight of 10 kD 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

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 high IgG titer and low IgM titer
• Fusion of one mouse’s spleen to the SP 2/0 myeloma cell line
• Screening of positive wells following fusion for IgG specific clones using immunizing antigen
• Cell line stabilization of up to 3 cell lines
• Isotype report on each final clone provided, confirming Ig class
• Cell line banking for 1 year following final clone shipment

Additional options and services include:

• Peptide synthesis (with antigen design assistance)
• Dual ELISA screening (i.e.: screen against positive hybridomas against GST or other large fusion partner) ($2400)
• Researcher screening of positive wells following fusion; 7-day screening time (starting at $500)
• Western blot or immunocytochemistry screening ($2200-$2600)
• Purchase of additional clones (from $600)
• Ascites production and purification
• Cell line isotyping ($50 per cell line)
• Cell line banking
• Cell bank generation

Downloads:

Monoclonal Antibody Development Packages (PDF) Monoclonal Antibody Development Order Form (PDF) Ascites Production Order Form (PDF) Terms & Conditions (PDF)