Total analytical solution for enabling robust mass spectrometric analysis of organoids and organ-on-chip samples

**During the project period, we have demonstrated significant progress in the analysis of organoids and organ-on-chip systems, which mimic the functions of human organs. These models provide researchers with unique opportunities to study diseases, test drugs, and develop new treatments without relying on animal models. We have addressed the complex requirements of analytical methods by developing and validating **AFFL (Automated Filtration/Filter Backflush)** – an innovative analytical platform.** Throughout the project, AFFL has been further developed into a system that combines advanced filtration and separation methods with sensitive mass spectrometers. This ensures precise and reliable measurements, even from small and complex sample volumes. We have successfully optimized the system to reduce variability in analyses, such as nonspecific binding during filtration, as well as improve sensitivity and accuracy. Additionally, we have fine-tuned key components such as columns, filtration technology, and mobile phases, which have contributed to improved analytical quality. This makes AFFL a comprehensive solution capable of addressing challenges that traditional methods have struggled to overcome. AFFL represents not only a technological solution but also a significant research methodology that paves the way for more precise and automated analysis. The results achieved during this work have laid the foundation for advancing biomedical research and drug development while simultaneously promoting the use of animal-free research methods.

The AFFL (Advanced Functional Flow Lab) platform supports the 3Rs (Replacement, Reduction, and Refinement) by enabling robust and precise analysis of organoid and organ-on-chip models, advancing their role as alternatives to animal testing. The platform addresses limitations in traditional methods by delivering high-quality results from small, complex sample volumes, promoting broader adoption of these advanced models. The project has enabled to elevate AFFL from a TRL 4 prototype to TRL 5-6 by developing consumable reagents and hardware. Collaborations with industrial partners, such as Merck Life Science, are being considered to accelerate market entry. AFFL has the potential to significantly enhance biomedical research, drug development, and the replacement of animal models.