Development of a High-Throughput Screening Platform to Study the Adsorption of Antigens onto Aluminum-Containing Adjuvants

Abstract

Aluminum-containing salts are important adjuvants in the formulations of many licensed human vaccines. However, in the early stage of the design of a new vaccine, a thorough understanding of the adsorption mechanisms of an antigen onto an aluminum salt is required. Therefore, we have developed a robust, rapid, and reproducible high-throughput screening (HTS) platform to study the adsorption capacity of aluminum-containing vaccines. The adsorption isotherms on aluminum hydroxide and aluminum phosphate of two model proteins, β-casein, and bovine serum albumin, were evaluated using a liquid handling system, which permitted rapid sample preparation in a small volume without nonspecific adsorption. Highly reproducible adsorption capacities and adsorptive coefficients were estimated based on the Langmuir model. To demonstrate the potential of this HTS platform, we evaluated the adsorption isotherms for two antigens, hepatitis B surface antigen and a pneumococcal serotype polysaccharide conjugated to a protein-D carrier, onto aluminum-containing vaccines at either a constant protein or a constant aluminum concentration. The automated assay enabled the rapid quantification of antigen adsorption with a significant reduction in operator workload and reagent use. This platform should accelerate data acquisition during the development of a new vaccine.