Monoclonal antibody (mAb) ascites fluid production in mice is a well described method of antibody production, although ethical questions regarding the pain and distress of the animals utilized in this process have been raised. In this study, mice were injected with pristane to initiate granuloma formation, followed by an injection of murine hybridoma PA 2II 2F9-1-1 (2F9) to produce IgG1 subclass mAb directed against protective antigen (PA) protein of Bacillus anthracis. Upon the recognition of pain or distress, characterized by well accepted clinical signs, analgesics were administered by treatment group. The control group (A) received saline, group (B) received meloxicam, group (C) received buprenorphine, and group (D) received both meloxicam and buprenorphine. Analgesics were administered by group for a total of 36–48 hours prior to the second ascites fluid collection. There was no statistical difference in the antibody titer or functionality between treatment groups at the first or the second collection time points. As reported here, analgesics may be administered upon recognition of pain in mice used for mAb ascites fluid production without affecting antibody concentration or quality and may warrant further evaluation as a refinement in other hybridoma cell lines. 1. Introduction Monoclonal antibody (mAb) ascites fluid production in mice is a well described method of antibody production [1–3]. mAb ascites fluid is typically produced in mice that are induced to form granulomas by intraperitoneal injection of pristane about fourteen days before injecting the hybridoma cell line. Pristane prevents the removal of hybridoma cells from the peritoneal cavity to the peripheral circulation, allowing growth of the hybridoma cells and the accumulation of ascites fluid containing the desired antibody [4, 5]. Ascites production is affected by several factors including the hybridoma cell line, timing of pristane injection in relation to inoculation of the hybridoma cells, and the age of mice [1, 6, 7]. The volume of pristane injected, the number of hybridoma cells injected, the frequency and method of abdominal paracentesis, and the hybridoma growth characteristics may all influence pain, distress, and survival [1, 6, 7]. Based upon observed clinical, pathophysiological, and pathological findings, the production of ascites is considered to cause pain and distress in mice [3, 6–8]. The injection of pristane induces a chronic inflammatory response in the development of granulomatous tissue and is not considered to cause pain in the short term [1, 8]. However, the
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