Quantitative Analysis of Vaccine Antigen Adsorption to Aluminum Adjuvant Using an Automated High Throughput Method.

Aluminum containing adjuvants have been widely used in vaccine formulations to safely and effectively potentiate the immune response. The examination of the extent of antigen adsorption to aluminum adjuvant is always evaluated during the development of aluminum adjuvant containing vaccines. A rapid automated high throughput assay was developed to measure antigen adsorption in a 96-well plate format using a TECAN Freedom EVO® (TECAN). The antigen adsorption levels at a constant adjuvant concentration for each sample were accurately measured at 12 antigen /adjuvant (w/w) formulation ratios. These measurements were done at aluminum adjuvant concentrations similar to normal vaccine formulations, unlike previous non-automated and automated adjuvant adsorption studies. Two high sensitivity analytical methods were used to detect the non-absorbed antigens. The antigen to adjuvant adsorption curves were fit to a simple Langmuir adsorption model for quantitatively analyzing the antigen to the adjuvant adsorption level and strength. The interaction of two model antigens, bovine serum albumin (BSA) and lysozyme (Lyso), with three types of aluminum adjuvant were quantitatively analyzed in this report. An automated high throughput methodologies combined with sensitive analytical methods is useful for accelerating practical vaccine formulation development.