Mouse Monoclonal Antibody Development Services

Hybridomas to Monoclonal Antibodies

Monoclonal antibodies are produced through fusion of murine B-cells (isolated from the spleen of immunized mice) with an immortal mouse myeloma cell line. Fusion of the two different cell types produces a "hybridoma" cell line that shares two desirable features of the parent cell lines: (1) secretion of monoclonal antibodies and (2) immortality. The resulting hybridoma cell line is an infinitely renewable source of monoclonal antibody. Development of mouse monoclonal antibodies through hybridoma fusion generates up to several hundred hybridoma clones secreting monoclonal (epitope-specific) antibodies against your target of interest. These monoclonal antibodies can then be further screened to identify clones with the desired characteristics.

Advantages of monoclonals include:

• Monoclonal antibodies bind to a single (defined) epitope on the target molecule and are routinely used in applications where binding specificity is a requirement.
• A variety of monoclonal antibodies can be generated against distinct epitopes on the target molecule in a single project.
• Screening of individual clones allows antibodies with the desired characteristics to be isolated.
• Monoclonal antibodies are secreted from a monoclonal hybridoma cell line that is immortal, thus providing a renewable source of the monoclonal antibody and ensuring lot to lot consistency.

Disadvantages of this approach include:

• More costly than polyclonal antibody development;
• Highly-conserved antigens may not evoke a strong immune response (due to tolerance). While a concern, this can be mitigated by using tolerance-impaired mouse strains such as NZB/W and through the use of immunogenic epitope tags/carrier proteins fused to the immunogen.

Custom antibody development is not a "one size fits all" process.

We pride ourselves on taking the time to learn your goals for the project and designing a project to meet those goals. "Begin with the end in mind" is a phrase that underlies everything we do, by starting with a solid plan we deliver solid results. Our experienced team can guide you through selection of: immunogen, immunization schedule, and hybridoma screening approach using our optimized platform. If you have particular requirements for your project (that differ from the overview below) we will work collaboratively with you to ensure the project meets your specifications.

Process Overview

Immunogen Selection (Variable timeline)

• We consult with you to determine the most appropriate antigen for development of the antibodies you require.

Immunization Schedule (6 weeks)

• Immunization of 5 mice.
• Accelerated schedule for production of high-affinity antibodies.

ELISA Titer Testing (Less than one week)

• ELISA testing of test bleeds to evaluate serum titer and select best-responding mouse for fusion and hybridoma creation.

Hybridoma Fusion and Screening (4-5 weeks)

• High efficiency electrofusion of spleen cells from best-responding mouse with mouse myeloma fusion partner; fusion plated across 25 x 96 well plates.
• High-throughput screening of hybridoma clones by ELISA (primary ELISA screen)
• Secondary ELISA screen on up to 94 candidate clones identified in primary screening.
• Cryo-preservation of up 94 candidate hybridoma clones for subcloning.
• Antibody-containing supernatants made available to you for internal testing if desired.

Sub-cloning of Selected Clones (6 weeks)

• Selected clones sub-cloned and cell-banked.
• Monoclonality confirmed by limiting dilution.

Related Services

Antibody production and purification from mouse monoclonal cell lines is also available. See Hybridoma mAb Scale-Up.