Monoclonal Antibody Ascites Fluid Production

Background
Monoclonal antibodies are exceptionally powerful research tools and also have potential clinical uses. Animals, usually mice, may be involved in two phases of the monoclonal antibody production process. The first stage involves sensitization of mice with the antigen, after which they are sacrificed to collect plasma cells. In the second stage, the plasma cells are used to produce hybridoma cells, which may be used in vitro to produce monoclonal antibodies, or they may be injected intraperitoneally into other mice, where they produce ascites tumors which yield an antibody-rich ascites fluid.

Priming
Pristane, the most commonly used priming agent for ascites production, is believed to act by inducing a glaucomatous reaction and by interfering with peritoneal fluid drainage. Priming with agents other than pristane must be specifically justified in the Animal Use Approval Form which is submitted to the IACUC for review.

Harvesting Ascites Fluid
Accumulation of fluid in the peritoneal cavity causes abdominal distention, discomfort and in extreme cases may even cause respiratory distress. The investigator must tap the excess fluid frequently enough to minimize this distention.

The animals will be observed at least three times per week for the first week after injection of the hybridoma cells. Afterwards, they will be observed daily. The ascites will be drained before it becomes excessive and before respiratory distress is evident.

Euthanasia
Animals bearing ascites tumors will not be maintained past a point where they are in good health. If an animal exhibits poor condition, debility, or discomfort (as evidenced by a hunched posture and ruffled hair coat), then it will be euthanized.