Digitally Enabled Generic Analytical Framework Accelerating the Pace of Liquid Chromatography Method Development for...

Area of Science:

• Analytical Chemistry
• Pharmaceutical Sciences
• Vaccinology

Background:

• The increasing complexity of vaccine formulations necessitates robust analytical methods for adjuvant characterization.
• Current methods for separating and quantifying vaccine adjuvants are time-consuming and labor-intensive, hindering rapid development.
• Reversed-phase liquid chromatography (RPLC) is crucial for vaccine quality control but faces challenges in method development speed.

Purpose of the Study:

• To introduce a generic, efficient analytical framework for the chromatographic resolution of common non-aluminum-based vaccine adjuvants.
• To enable faster and more proactive assay development for complex adjuvant-antigen mixtures in vaccine formulations.
• To establish a versatile approach for method development using in silico simulations and a dynamic retention time database.

Main Methods:

• Extensive stationary phase screening and multifactorial in silico simulations of adjuvant retention times.
• Development of retention models based on gradient time, temperature, organic modifier blending, and buffer concentration.
• Utilized LC with diode array detector (DAD), charged aerosol detector (CAD), and mass spectrometry (MS) for comprehensive analysis.

Main Results:

• Generated 3D resolution maps enabling excellent baseline separation of multiple adjuvants in a single run.
• Achieved high accuracy in simulations, with experimental and simulated retention time differences below 1%.
• Demonstrated the framework's capability for rapid generation of analytical assays (content, purity, identification) for complex vaccine formulations.

Conclusions:

• The developed analytical framework significantly accelerates assay development for vaccine adjuvants.
• The in silico approach and dynamic database offer a versatile and proactive solution for complex mixture analysis.
• This method supports the development of novel adjuvant formulations by providing accurate and precise quantitative data.