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) that the core staff doesn't immunize.

Three to five BALB/c mice are immunized for each antigen. For each project, investigators must provide a minimum of 5 mg of peptide coupled to keyhole-limpet hemocyanin (KLH) carrier protein and 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 weeks to 10 weeks. Once the mice are immunized and boosted, individual bleedings are performed. From each mouse, approximately 100 µl each of pre-immune and immune serum is collected and aliquoted into 20-µl samples. These samples are provided to the investigator, who may then test them for the presence of the relevant specific antibody. The core also can perform ELISA screening of the serum.