Four-step approach to efficiently develop capillary gel electrophoresis methods for viral vaccine protein analysis

Introduction: Vaccines against infectious diseases are urgently needed. Therefore, modern analytical method development should be as efficient as possible to speed up vaccine development.
Objective: The critical method parameters (CMPs) were to be identified and a set of steps to efficiently develop and validate a CE-SDS method for vaccine protein analysis based on a commercially available gel buffer was to be established.
Methods: CMPs were obtained from reviewing literature and testing the effects of gel buffer dilution. A four-step approach, including two multivariate DoE steps, was proposed based on CMPs and was verified by CE-SDS method development for (i) the determination of influenza group 1 mini-haemagglutinin glycoprotein, and (ii) the determination of polio virus particle proteins from an inactivated polio vaccine (IPV).
Results: CMPs for sample preparation were incubation temperature(s) and time(s), pH, and reagent(s) concentration(s), for detection the detection wavelength. The effects of gel buffer dilution revealed the CMPs for CE-SDS separation to be the effective length, the gel buffer concentration, and the capillary temperature. The four-step approach based on the CMPs was efficient for the development of the two CE-methods.
Conclusion: A Four-step approach to efficiently develop capillary gel electrophoresis methods for viral vaccine protein analysis was successfully established.