Title : Fluorescence A-TEEM Method for Vaccines Characterization
Abstract:
We will present several examples to highlight the capabilities of the fluorescence A-TEEM method, a novel approach which combines UV/Vis with fluorescence EEM spectroscopy. The benefit of this approach is the ability to provide robust and rapid spectroscopic characterization of vaccine components and vaccine formulations, samples that are extremely challenging to traditional spectroscopies, such as Raman and NIR. The Coronavirus pandemic applied historic pressure on vaccine development and production, collapsing development timelines from years to months, with many firsts in formulation and production. Requirements for product quality still had to be met though, highlighting the need for rapid analytical techniques to characterize vaccines from R&D to formulation development, and through manufacturing to final QA/QC. Spectroscopic techniques are known to be rapid and are therefore used extensively for PAT and QA/QC testing. However, the “go-to” techniques such as Raman and NIR often do not work for vaccines, as they struggle with low protein concentrations typical of these formulations. The fluorescence A-TEEM method is a unique alternative, combines high sensitivity and specificity, with limits of detection to 0.015 ug/mL, and data acquisition times typically under 60 seconds. As the method combines two well established techniques (UV/Vis and fluorescence), the technique can be validated by following the relevant USP chapters, <853> and <857>. We will present results from an A-TEEM study on four closely related combination vaccine formulations, Solo-Jec brand canine vaccines from Boehringer Ingelheim VetMedica. A-TEEM was able to accurately differentiate between the four formulations, even when they differed by only a single coronavirus component. To ensure repeatability, calibration data (two separate samples for each formulation) were collected, and reproducibility was assessed with a third unique set of validation samples, collected on a different instrument by a different operator. The A-TEEM is able to identify and validate “unknown” samples with 100% certainty. In addition to vaccine formulations, we will present AAV characterization studies, where the A-TEEM was able to rapidly (<90sec) differentiate biotinylated AAV samples that are complexed with streptavidin-dye conjugates from uncomplexed AAVs.
