Rat Monoclonal Antibodies: Custom Development

Rat monoclonal antibodies offer several advantages over murine antibodies. We use the same murine myeloma partner as our murine hybridoma process so the resulting hybridomas are chimeras. Not only does the rat host allow the development of monoclonal antibodies to murine proteins but rat antibodies pair well with murine monoclonals in sandwich assays. With two different species of antibodies, there is no cross reaction of available anti-mouse secondary antibody binding to the capture antibody.

Advantages

  • Used for murine antigens.
  • Reduced antigen requirement. Single rat immunized versus multiple mice.
  • Host variation offers different antibody responses.
  • Larger spleens yield twice the number of hybridomas per fusion.
  • No special growth media required.
  • Purification methods similar for murine antibodies
  • Pair with existing mouse antibodies in sandwich assays
 

Schedule for a typical rat hybridoma project:

Phase I: Immunization (4-6 weeks)

Approximately 0.5-1.5 mg of protein is required for standard immunizations. A serum titer will determine when we schedule a fusion.

Phase II: Fusion (2 weeks from fusion to screening)

At this stage, splenocytes are plated into either 32 or 64 x 96 well plates, and screened by solid-phase ELISA for antigen specific clones. Fusion positives are expanded into 24-well plates and can be rescreened using a number of different assays that will help you identify the most desirable clones for your needs.

Phase III: Subcloning (2 weeks)

Fusion positive clones are subcloned by limiting dilution and screened for antigen specific positive clones by solid-phase ELISA.

Phase IV: Characterization and purification

We offer a wide variety of services including antibody production and purification, antibody characterization, and cell line archival. Please contact us for more information.

Additional Services