Determination of percentage antigen adsorption in vaccine drug formulation.

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Production of vaccine drug formulation is a multi-million dollar commercial activity. To operate on an industrial scale normally requires the manufacturing of high efficiency drug products which is typically obtained by the addition of adjuvants in drug formulations, but percentage adsorption of antigens vary dramatically among different drug formulations. Since immunogenicity is directly linked with percentage antigen adsorption, it is extremely critical to develop a method for the quantification of adsorbed antigens in the final drug product. Determination of percentage antigen adsorption would require the quantification of both free and adsorbed antigens in the drug product. Sanofi Pasteur is attempting to develop a method for quantification of free antigens using AQUA method coupled with LC-MS. The method is currently being optimized to overcome the matrix effect. The quantification of adsorbed antigens is still a challenge and we propose to desorb antigens using various reagents and temperature conditions to breakdown the strong electrostatic interactions between antigens and adjuvant. The drug product will be centrifuged to isolate free antigen. The pellet containing adsorbed antigen will be resuspended and treated with reagents such as cetylpyridinium chloride, sodium citrate dihydrate, sodium phosphate dibasic, guanidine HCl, urea and succinate. Once the antigens are successfully desorbed, the free antigens will be digested with trypsin and quantified by LC-MS method being developed by Sanofi Pasteur. This method will be an advancement in pharmaceutical research that will enhance drug efficacy in patients and bring increased visibility and recognition to Canadian research within the world-wide scientific community.