Behavioral, pharmacokinetic, and B cell comparison of alum and MF59. BALB/c mice were immunized with oxycodone vaccine OXY-KLH plus alum, OXY-KLH plus MF59, or a KLH control. (a) Mice were immunized on days 0, 14, 28, and 56, and OXY-specific IgG titers at 59 days were measured by ELISA (n = 7 per group). (b-c) Mice were immunized on days 0, 14 and 28, and motor activity induced by 5 mg/kg oxycodone was recorded over 60 minutes post-injection on day 45 (n = 7 per group). (c) Cumulative motor activity as total distance travelled during the first 15 minutes post-injection. (d) Mice were immunized on days 0, 14 and 28 and challenged with 2.25 mg/kg oxycodone on day 35, and oxycodone concentrations in brain 30 minutes after oxycodone administration were measured by GC-MS. (e) Mice were immunized on days 0, 14 and 28, and lymph nodes and spleens were collected at 14 or 42 days post-immunization (n = 5 per group). Total OXY-specific B cells populations were analyzed by flow cytometry. Data are mean±SEM. *p < 0.05, **p < 0.001, **** p < 0.0001.