Purification and quantitative description of Rhodococcus equi IgG designed for aerosol nebulization to foals

The objective of this study was to purify IgG from commercially available hyperimmune Rhodococcus equi plasma and to assess the delivery of IgG as an aerosol to the equine lung. IgG was purified from plasma, and the IgG concentration of both the plasma and the purified IgG was determined by ELISA. The purified IgG was aerosolized using a vibrating mesh nebulizer and aerosol characterization was performed using cascade impaction. The purified IgG was nebulized to six healthy adult horses in order to assess the efficacy of pulmonary delivery and safety of administration. Bronchoalveolar fluid was retrieved endoscopically using a low volume technique prior to aerosolization (time 0) and at 0.5, 4 and 24 hours post aerosolization. The BAL fluid IgG concentration was determined and cytologic analysis was performed.

The IgG concentrations of the plasma and purified IgG were 2,175 mg/dL and 1,145 mg/dL, respectively. The MMAD of the purified IgG aerosol was 4.7 microns. The mean BAL fluid IgG concentration increased 61% from 19.33 µg/dL at time 0 to 31.5 µg/dL at 0.5 hours, but this increase was not significant (P=0.603). No significant change was observed in inflammatory cell numbers over time or at any time point during the study. This study demonstrated that IgG antibodies were purified at a concentration acceptable for nebulization, and that the nebulization unit generated aerosol particles from the IgG solution of appropriate size for pulmonary delivery. Nebulization of purified IgG to adult horses was well tolerated and caused no local or systemic adverse effects.