Analytical Quality by Design Method Development for Vaccine Characterization

Sammanfattning: Vaccines that are safe, efficacious, and can be rapidly developed are needed to prevent and to react to emerging global infectious disease threats such as influenza, Polio, and Coronavirus diseases. Fast and reliable analytical methods are required without delay to support vaccine process and product development, characterization, and quality control testing. The traditional analytical methods for vaccines are laborious and often lack analytical power, causing slow, expensive, or sometimes failing vaccine development. Capillary electrophoresis (CE) is a technique that has great potential for biopharmaceutical analysis, although there has been limited application in vaccine development.Several novel CE methods were explored, developed, and applied for viral vaccine analysis, making use of the analytical quality by design (AQbD) process and tools. AQbD is a framework of science- and risk-based decision making to achieve in-depth method understanding and to set up fit-for-purpose and in-control analytical methods. Commercial kits for capillary gel electrophoresis (CGE) and imaging capillary isoelectric focusing (icIEF) for antibodies analysis were applied and improved for vaccine analysis. Analytical mechanisms were studied, such as the effect of gel buffer composition on separation, and an AQbD CGE method development strategy was established. The strategy was successfully applied to develop CGE methods for the analysis of seasonal and universal influenza, and sabin inactivated polio vaccine proteins. An icIEF method was also developed, validated, and applied for the universal influenza vaccine protein. A capillary zone electrophoresis (CZE) method development for intact adenovirus concentration determination started with background electrolyte (BGE) and capillary design and screening. An BGE with tris and tricine and a neutral capillary resulted in optimal and robust separation and limited adsorption. The CZE method was validated for seed release, in-process control, product release, and stability testing. The precise, accurate, fast, and robust CZE method was applied for all process intermediates and used at different locations. Process impurities and product degradation could also be characterized.Additionally, CZE methods for chloride and bromide analysis in complex matrices, and a CGE method for host cell DNA characterization were developed for characterization as well as to support process development.Development of CE methods using AQbD reduced lead times and costs. The developed CE methods were easier to use, were more accurate and precise, and were more selective for product and process impurities compared to the previously used analytical methods for vaccines. The use of CE and AQbD helped improve on vaccine safety, efficacy, and quality.