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IBEC Seminar: Miguel Holgado
viernes, abril 1, 2022 @ 10:00 am–1:00 pm
Technologies for highly multiplexed in-vitro diagnostic systems and organ on chips. Cases studies for COVID-19 and neurodegenerative diseases
Miguel Holgado, Centro de Tecnología Biomédica-Universidad Politécnica de Madrid and Instituto de Investigación Sanitaria del Hospital Clínico San Carlos
Technologies for biomarkers screening are of very high importance, particularly when they are reliable, cost effective, easy to use for measuring multiples biomarkers in a single diagnostic KIT working with real biological samples such as serum, saliva, wastewater or culture medium. In this paper we report technologies for the development of a highly multiplexed kit for detecting biomarkers of COVID19 in serum, saliva and wastewater analyzing their correlation with the severity of the COVID19 and showing relevant figures about the severity (90 patients in serum), immunity (200 volunteers in saliva donating sample every 10 days in three months), and wastewater. This technology has also demonstrated for measuring particular proteins of the SARS-COV-2 in wastewater, which results have been compared with Polymerase Chain Reaction. Recently we are using these technologies for detecting Alzheimer Disease (AD) biomarkers in serum.
In concrete, we have achieved to detect the total tau protein at the level of 10 pg mL-1 in serum as a biomarker for early detection of the AD. Other biomarkers are also under development right now. In this term, the use of advanced in vitro diagnostic systems with organ-on-chip based technologies are of a high relevance because can be used for monitoring relevant biomarkers secreted by the cells, tissues or biopsies in these types of bioreactors. We have recently developed microfluidic chips acting as bioreactors for neuronal circuits on a chip for biological network monitoring and brain slice-on-a-chip for organotypic culture and effective fluorescence injection testing. Cultured neuronal networks (CNNs) are a robust model to closely investigate neuronal circuits’ formation and monitor their structural properties evolution. Typically, neurons are cultured in plastic plates or, more recently, in microfluidic platforms with potentially a wide variety of neuroscience applications. As a biological protocol, cell culture integration with a microfluidic system provides benefits such as accurate control of cell seeding area, culture medium renewal, or lower exposure to contamination. In this paper it is presented a novel neuronal network on a chip device, including a chamber, fabricated from PDMS, vinyl and glass connected to a microfluidic platform to perfuse the continuous flow of culture medium.
As a step forward, we employ this technology as an alternative brain slice-on-a-chip, integrating an injection system inside the chip to dispense a fluorescent dye for long-term monitoring. Hippocampal slices are cultured inside these chips, observing fluorescence signals from living cells, maintaining the cytoarchitecture of the slices. Having fluorescence images of biological samples inside the chip demonstrates the effectiveness of the staining process using the injection method avoiding leaks or biological contamination. The technology developed in this study presents a significant improvement in the local administration of reagents within a brain slice-on-a-chip system, which could be a suitable option for organotypic cultures in a microfluidic chip acting as a highly effective bioreactor.
 Developing an Optical Interferometric Detection Method based biosensor for detecting specific SARS-CoV-2 immunoglobulins in Serum and Saliva, and their corresponding ELISA correlation. Sensors & Actuators: B. Chemical 345 (2021)
 A new optical interferometric in-vitro detection for Alzheimer´ disease diagnostic in Serum. To be published elsewhere.
 Neural circuits on a chip for biological Network Monitoring. Biotechnology Journal 2021. https://doi.org/10.1002/biot.202000355
 Alternative Brain Slice-on-a-Chip for Organotypic Culture and Effective Fluorescence Injection Testing. Int. J. Mol. Sci. 2022, 23, 2549.