Optimization of Anti-Rh D Immunoglobulin Stability in the Lyophilization Processes

Objective Anti-Rh D IgG is used for the prevention of anti-D antibody production in Rh- individuals who have been exposed to Rh+ red blood cells. The stability of IgG preparations as a solution is low, with a shelf life of a year or more. Formulation of anti-Rh D IgG as a lyophilized preparation would decrease its degradation rate and increases its shelf life. The objective of this study was to formulate the anti-Rh D as a lyophilized preparation using different formulations and optimize the lyophilization processes. Materials and Methods The effect of various formulations on the stability of anti-Rh D was evaluated using accelerated stability test.  In this method the amount of transmittance (T %) at ۵۸۵ nm for the lyophilized preparations had inverse relationship with aggregation of anti-Rh D. To improve stability, the most stable formulation was selected and different concentrations of sucrose in the presence of sodium-potassium phosphate buffer ۲۵ mM pH ۷.۵. Then, the bioactivity was determined, using the ELAT test and also, the amount of moisture measured in this formulation. Results Among different formulations, the one with anti-Rh D ۵ mg/ml, tween ۸۰ ۰.۱%, glycine ۰.۱۵ M, manitol ۷% and sucrose ۶۰ mM in sodium-potassium phosphate buffer ۲۵ mM pH ۷.۵ was the most stable formulation (P<۰.۰۵). The result of biological test of ELAT showed that bioactivity of more than ۹۳% meets the requirement set by British Pharmacopoeia. The amount of moisture measured in this formulation was less than ۳%.    Conclusion It was concluded that this formulation could be introduced as a candidate for the formulation of anti-Rh D in a lyophilized dosage form.

Keywords:

Anti-Rh D IgG , Bioactivity , Formulation , Lyophilization , Protein stability