Pharmacokinetic sampling and assays
Pharmacokinetic blood samples were drawn immediately pre-dose and at 0.5, 2, 4, 6, and 24 h, then 7 days, following the first dose, and 28 and 70 days after the final dose. Serum was separated from blood and stored at −70°C prior to analysis for MOv18 IgE concentrations by indirect ELISA, using a fully validated assay. Briefly, MOv18 IgE antibodies were captured by recombinant FRα (BioTechne 5646-FR) and detected by an anti-IgE-HRP detection antibody (Sigma A9667). Levels were quantified by a MOv18 IgE standard curve with a limit of quantification of 2.5 ng/ml and a dynamic range of 2.5–200 ng/mL in serum. The following pharmacokinetic parameters were evaluated: terminal half-life calculated from the terminal slope of the log concentration–time curve (