Immunization and Antiserum Collection
Though it's possible to produce monoclonal antibodies using mice, rats and rabbits, the Antibody Hybridoma Core only immunizes mice. When immunized, boosted and bled by core staff members, these mice will be housed in a pathogen-free colony. Investigators are responsible for housing and maintaining animals (mice, rats and rabbits) not immunized by core staff.
Three to five Balb/c mice are immunized for each antigen. For each project, investigators must provide a minimum of 5 milligrams (mg) of peptide coupled to keyhole-limpet hemocyanin (KLH) carrier protein, as well as 5 mg of free peptide. The amount of protein required will vary, though 2 mg is optimal.
The timeline for immunizing and boosting can range from four to 10 weeks. Once the mice are immunized and boosted, individual bleedings are performed. From each mouse, approximately 100 microliters each of pre-immune and immune serum is collected and aliquoted into 20-microliter samples. These samples are provided to the investigator, who may then test them for the presence of the relevant specific antibody. The core is also able to perform enzyme-linked immunosorbent assay (ELISA) screening of the serum.