Background: The growing demand for versatile laboratory automation is exemplified in the context of liquid biopsy, where multi-analyte approaches are increasingly recognised for their potential to enhance diagnostic sensitivity in oncology. However, current practice often necessitates the use of dedicated instruments and workflows for the extraction of each analyte, posing financial and logistical barriers for automated multi-analyte liquid biopsy. Methods: Here, we present Robotic Centrifugal Microfluidics (RoCM), an all-in-one platform that combines the versatility of centrifugal microfluidics and operational flexibility of robotic liquid handling. This combination enables the automation of complex micro- and macrofluidic protocols, realised through the use of (1). exchangeable microfluidic cartridges and (2). programmable robotic operations such as in-rotation liquid supply, magnetic bead manipulation, or microfluidic valving. In-rotation robotic liquid manipulation maintains fluid control under centrifugal forces and reduces the cartridge footprint associated with pre-loaded liquid reservoirs. Platform applicability was demonstrated using two exemplary liquid biopsy workflows: the extraction of cell-free DNA (cfDNA) from blood plasma using RoCM-cfDNA slices and the extraction of extracellular vesicles (EVs) from blood plasma using RoCM-EV slices. Results: In a pilot study with patient samples from different cancer entities, the RoCM-cfDNA slices yielded comparable variant allele frequencies to a commercial bead-based instrument, while the RoCM-EV slices achieved a recovery of a greater diversity of EV subpopulations than semi-automated size-exclusion chromatography. Conclusions: By simply exchanging cartridges, RoCM enables the extraction of diverse analytes within a single automated system. Its application can be extended to further analytes, such as circulating tumour cells (CTCs), or to applications beyond liquid biopsies, where versatile micro- and macrofluidic protocols benefit from implementation in a single automation instrument.

Food allergies pose a serious and increasing public health challenge as many allergens can trigger potential life-threatening reactions in affected individuals. Reliable detection of allergens such as soy proteins is therefore a critical and essential tool to detect cross-contaminations, particularly in food processing and production environments. To enable monitoring directly at these sites, portable biosensing platforms are required to provide rapid point-of-need detection. However, the development of such systems requires the combination of biochemistry, material science, and precision engineering. In this article we present a centrifugal microfluidic platform integrating automated sample preparation, clean-up, and immunological detection in a single-use disposable cartridge. Targeting soybean protein as a model allergen, the system combines optimized immunoassay chemistry with advanced polymer microfabrication. Regioselective antibody coupling strategies were investigated for stable and oriented bead conjugation. N-glycan click chemistry was used to produce monodisperse, highly functional fluorescent microbeads. The microfluidic LabDisk was first manufactured by thermoforming and later transferred to injection molding with integrated optical elements to increase sensitivity. The fully integrated LabDisk detected soy protein at the lowest concentrations tested, namely 1 ppm in extraction buffer and 4 ppm in incurred complex matrices. This work shows the important steps from biochemical concept to scalable manufacturing, paving the way for mass-produced, portable allergen biosensors.

Background: The growing demand for versatile laboratory automation is exemplified in the context of liquid biopsy, where multi-analyte approaches are increasingly recognised for their potential to enhance diagnostic sensitivity in oncology. However, current practice often necessitates the use of dedicated instruments and workflows for the extraction of each analyte, posing financial and logistical barriers for automated multi-analyte liquid biopsy. Methods: Here, we present Robotic Centrifugal Microfluidics (RoCM), an all-in-one platform that combines the versatility of centrifugal microfluidics and operational flexibility of robotic liquid handling. This combination enables the automation of complex micro- and macrofluidic protocols, realised through the use of (1.) exchangeable microfluidic cartridges and (2.) programmable robotic operations such as in-rotation liquid supply, magnetic bead manipulation, or microfluidic valving. In-rotation robotic liquid manipulation maintains fluid control under centrifugal forces and reduces the cartridge footprint associated with pre-loaded liquid reservoirs. Platform applicability was demonstrated using two exemplary liquid biopsy workflows: the extraction of cell-free DNA (cfDNA) from blood plasma using RoCM-cfDNA slices and the extraction of extracellular vesicles (EVs) from blood plasma using RoCM-EV slices. Results: In a pilot study with patient samples from different cancer entities, the RoCM-cfDNA slices yielded comparable variant allele frequencies to a commercial bead-based instrument, while the RoCM-EV slices achieved a recovery of a greater diversity of EV subpopulations and up to one magnitude higher recovery than semi-automated size exclusion chromatography. Conclusion: By simply exchanging cartridges, RoCM enables the extraction of diverse analytes within a single automated system. Its application can be extended to further analytes, such as circulating tumor cells (CTCs), or to applications beyond liquid biopsies, where versatile micro- and macrofluidic protocols benefit from implementation in a single automation instrument.

Food allergies are a severe burden for affected individuals and healthcare systems. To tackle the need for simple food allergen detection, we developed a system for the detection of the soy protein glycinin via a centrifugal microfluidics-assisted lateral flow immunoassay (LFIA). Glycinin is a complex allergen requiring extensive sample preparation. The presented workflow includes a manual denaturing extraction, followed by automated centrifugal microfluidic desalting, metering and detection via LFIA. The functionality of the microfluidic cassettes was tested on prototypes produced via microthermoforming before an injection molding tool was designed, which added a cylindrical lens to improve the readout. Overall, this system aims to aid in food allergen detection with high sensitivity and minimized manual steps.