Multivariate approach for screening and optimization of RP-UPLC method performance for a formulated vaccine

The application of QbD principles to analytical method development is focused on the concept to build quality during method development. for this reason, the methods to be developed had to satisfy requirements described in Analytical Target profile (ATP). In particular, the resulting methods have to satisfy: I) minimum requirements related to method performance (precision, sensitivity, accuracy, selectivity etc.) and II) minimum requirements related to assay feasibility that is applicability to a routine Quality Control testing. Recombinant proteins are the active components of the Meningococcal Vaccine and the control of method variables is important to assure the RP-UPLC assay quality. For this reason, preliminary screening studies of two critical factors, identified in vials type and antigens concentration, were performed to select the best operative conditions and ensure sensibility and repeatabaility of the method. To achieve this goal, asymmetric screening matrices (based on Free-Wilson model) were set up to study the two factors. The software used for experimental plans construction was NEMRODW(R) 2015. The graphic effects study of the single variables and their interactions allowed identifying the optimum operative conditions.