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Networking Buffet Lunch -- Network with Exhibitors and View Posters

Mid-Afternoon Coffee Break and Networking

Best Poster Awards Sponsored by Lab-on-a-Chip Journal, Royal Society of Chemistry (RSC) -- In the Coolsingel Room

Tackling the Global Health Threat Posed by Fungal and Oomycete Diseases Using Lab-on-a-Chip Devices
Ayelen Tayagui, Research Fellow, University of Canterbury, New Zealand

Lab-on-a-Chip devices can enhance the study of disease-causing microorganisms and tackle their effects on global health.  We describe the design, manufacture and use of devices use to study the forces these organisms use to to infect their hosts.

Towards Continuous Cytokine Monitoring in Organ-based Platforms
Maud Linssen, Researcher, Eindhoven University of Technology (TU/E), Netherlands

Investigations on organ-based platforms such as organoids, organ-on-a-chip, and transplantation organs, require monitoring strategies in order to optimally control the biological systems. However, there is a lack of tools to continuously measure specific low-concentration biomarkers with minimal perturbation. Biosensing by Particle Motion (BPM) is a sensing method with single-molecule resolution that has been specifically designed to enable the continuous monitoring of biomolecules at low concentrations, such as nucleic acids, metabolites, proteins and hormones. The method relies on tracking the motion of individual biofunctionalized particles (1 µm in diameter) that interact with a biofunctionalized substrate. The particles switch between bound and unbound states due to reversible single-molecule interactions, dependent on the presence of analyte molecules. Recently, sampling by microdialysis has been investigated [3]. In this paper we present the development of a BPM sensor to measure cytokines, exemplified with the detection of Interleukin-6 and TNF-a. We will present a study of molecular binders and coupling strategies, focusing on sensitivity and reversibility of the sensor. Furthermore, measurements with microdialysis will be shown. Finally, we will discuss the prospects of using BPM and microdialysis for the continuous monitoring of low-concentration biomarkers in organ-based platforms.

ElectroMed – A Programmable Peptide/Protein Screening Platform for Immunology
Cesar Pascual García, Lead Research Scientist, Luxembourg Institute of Science and Technology

Immunology has advanced significantly in recent years thanks to a better knowledge of molecular protein recognition, which has allowed for the creation of innovative treatment options such as genetic vaccines and cell therapies. However, there is a need to accelerate the process of epitope recognition by proteins in order to generate personalized therapies. ElectroMed has been a project with six European partners in which we created a concept for a programmable peptide screening platform based on an array of Immuno-FET sensors with programmable peptide receptors synthesized in-situ using electrochemically generated acid.

A proof-of-concept has been demonstrated by controlling the deprotection of acid labile groups, which enables a high purity synthesis of crude peptides. We remove diffusion-limited processes in chemistry by automating the peptide synthesis and using microfluidic flow in-situ synthesis in planar sensors, allowing us to expedite time to results. FET sensors have the potential to provide online quality control of peptide synthesis by detecting the fingerprints of acid labile group protection and deprotection, as well as superior information quality because label-free sensors can provide kinetic information.

In conclusion, our approach has the potential to speed research into peptide protein screening of linear peptide sequences. In this talk we will review our results and provide the context for the applications in immunology.

Parallel High-Throughput Single-Cell Printing Platform for Optoporation-Mediated Large Cargo Delivery
Gayathri R, PhD Scholar (Student), Indian Institute of Technology (IIT)-Madras, India

Effective intracellular delivery of biomolecules is crucial in analysing and engineering cell functions, for applications in therapeutic development, diagnostics, and drug delivery towards personalised medicine. Traditional bulk cell culture models employed so far for intracellular delivery, overlook cell population heterogeneity, while single-cell patterning offers robust statistical analyses without compromising cellular variability. In this work, a high-throughput single-cell patterning approach using microcontact printing is employed, where a micro-pillar stamp is used to imprint distinct proteins and finally pattern single-cell to small clusters of cells depending on the micro-pillar diameters. This approach is universal for any protein-cell combination and has 97-99 % patterning efficiency. This single-cell patterning was used as a platform for massively parallel optoporation-mediated intracellular delivery of small to very large biomolecules such as PI dye (668 Da), Dextran (3000 MW), SiRNA (20-24 bp), and enzyme (464 kDa). An easy-to-fabricate and simple-to-use 2D array of titanium micro-dish device is used to facilitate near infrared mediated optoporation. which disrupts the cell plasma membrane, allowing biomolecules to enter into cells with high efficiency (~96±2%) and cell viability (~98±1%). Our compact, robust, and simple-to-print approach has potential for single-cell therapy and diagnostics research, providing high specificity and subcellular accuracy in single-cell level biological processes.

Temporal Influence of Extracellular Vesicle-Depleted Serum on Extracellular Vesicles and Endothelial Cells
Luiz Fernando Cardoso Garcia, Scientist, Biociências e Biotecnologia do Instituto Carlos Chagas, Brazil

Extracellular vesicles (EVs) are generated by various cell types, including endothelial cells. The presence of EVs in fetal bovine serum (FBS), commonly used in cell culture, has been recognized as a potential confounding factor. To elucidate this, human brain microvascular endothelial cells (HBMEC) were cultured for 2 or 24 hours in the presence of EV-depleted FBS (EVdS). Cell viability, gene and protein expression, and EVs isolated from these cells were assessed. Additionally, EV uptake, ICAM-1 expression, and monocyte adhesion to HBMEC exposed to EVs were also examined. Elevated apoptosis rates in cells cultured with EVdS for 2 and 24 hours was observed. Within 2 hours, there was an upregulation of IL8, followed by the downregulation of IL6 and IL8 after 24 hours. Proteomic analysis revealed that EVs cultured for 2 hours (EV2h) were enriched in proteins associated with ribosomes and carbon metabolism, while those cultured for 24 hours (EV24h) exhibited proteins linked to cell adhesion and platelet activation. Moreover, HBMECs exposed to EV2h displayed increased ICAM-1 expression and monocyte adhesion compared to cells exposed to EV24h. These findings highlight that HBMECs cultured with EVdS produce EVs with distinct physical characteristics and protein content that varies across time.

Optical Bunching of Crystals in a 3D Printed Microfluidic Device for Enhancing Sample Delivery in Serial Crystallography
Anusha Keloth, Postdoctoral Research Fellow, Deutsches Elektronen-Synchrotron DESY, Germany

This paper presents the optical bunching of protein microcrystals in a 3D printed microfluidic device for improving the sample delivery for serial crystallography studies at X-ray free electron lasers.

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