Point-of-Care, Biosensors & Mobile Diagnostics Europe 2019 Agenda

Conference Registration, Materials Pick-Up, Morning Coffee and Tea in the Exhibit Hall

Morning Coffee, Tea and Networking in the Exhibit Hall

Networking Lunch in the Exhibit Hall and Poster Viewing

Commercialization of Microfluidic Devices for Point-of-Care Applications
Vincent Linder, Founder and President, CDP BioMedical Consulting and CEO at Calciscon AG, Portugal

This presentation will focus on the commercialization challenges specific to microfluidic devices for point-of-care applications.  At an early stage of development, strategies can focus on de-risking analytical and clinical performances and may overlook other critical aspects of the design.  For a successful commercialization effort, it is essential to implement at the onset of the program a comprehensive vision encompassing the patient presenting in a POC setting, the POC user and all the steps needed to obtain an actionable test result.  This presentation will focus on selected commercialization challenges and discuss directions/solutions to overcome them.

Bio-Inspired Laser-Induced Electrochemical Sensing Strategies: Antigen/Nanobody-based Magnetosensing
Karolien De Wael, Full Professor, Antwerp University, Belgium

In my talk a sensing strategy inspired by the advantages of enzymes and photoelectrochemical sensing, namely the integration of aerobic photocatalysis and electrochemical analysis, will be presented. The photosensitizer, a bioinspired perfluorinated Zn phthalocyanine, generates singlet-oxygen from air under visible light illumination and oxidizes analytes, yielding electrochemically-detectable products while resisting the oxidizing species it produces. Compared with enzymatic detection methods, the proposed strategy uses air instead of internally added reactive reagents, features intrinsic baseline correction via on/off light switching and shows C-F bonds-type enhanced stability. It also affords selectivity imparted by the catalytic process and nano-level detection, such as 20nM amoxicillin in ml sample volumes.

Additionally, this strategy can be applied to improve the sensitivity of assays by using magnetic beads in combination with an amperometric readout in a configuration so-called electrochemical magnetosensor. The main difference between of a magnetosensor from a biosensor is that a biorecognition element is linked to magnetic beads and can be temporally dispersed in a sample for improving binding kinetics and, then, accumulated on an electrode by a magnet to register an analytical response. In comparison with conventional optical read-out systems, electrochemical sensors can register an analytical response almost immediately and without the need of developing the reaction in bulk volume, because the response of electrochemical sensors is determined by a thin layer of measuring solution near the electrode surface where the beads with immunoreagents are placed. This approach may improve sensitivity and decrease analysis time in comparison to conventional ELISA.

Label-Free Biosensing With Impedimetric and Thermal Detection Methods
Patrick Wagner, Professor, Soft-Matter Physics and Biophysics Section, Departement Natuurkunde en Sterrenkunde, Katholieke Universiteit Leuven, Belgium

The first part of this presentation addresses the detection of single-nucleotide polymorphisms in DNA by monitoring the denaturation of double-stranded DNA fragments on solid chips such as synthetic diamond coatings. Denaturation experiments were performed in two different ways: i) Chemically, with NaOH solutions in combination with impedance spectroscopy as readout principle and ii) thermally, using the heat-transfer method HTM. The latter technique measures the thermal interface resistance Rth between the chip and the liquid; the Rth parameter responds sensitively to all molecular changes at the solid-liquid interface.

Using these two methods, I will address a couple of “real life” biosensing applications such as the detection of the allergen Ara h1 (a protein) in peanut butter, the neurotransmitter serotonin in blood, and histamine in fish brine and intestinal fluids. Histamine is a mediator in the irritable bowel syndrome IBS and we are currently working towards a catheter-based diagnostic tool. Within these applications, polymer-based receptors (molecularly imprinted polymers, MIPs) are playing an advantageous role owing to their long-term stability, chemical resilience under adverse conditions, and their regeneration capacity.   

In the final part of the presentation, I will discuss the selective detection and identification of cells (e.g. macrophages, cancer cells) and bacteria using whole-cell receptors. These receptors, surface-imprinted polymers (SIPs), are fabricated by soft lithography with stamping of template cells onto polyurethane layers. Special attention goes to the rebinding mechanism between the imprints and target cells and to ways how to boost selectivity by repetitive exposure. Combining these SIP receptors with HTM readout, relevantly low detection limits are becoming feasible. Especially for agriculture, the food industry and in the context of environmental safety, there is a considerable need for fast, on-site techniques to detect pathogenic microorganisms.

Afternoon Coffee and Tea Break and Networking in the Exhibit Hall

Rapid Diagnosis of Breast Cancer: Innovative Approaches with a Focus on Low Resource Setting
Jane Brock, Chief of Breast Pathology, Brigham and Women’s Hospital, Harvard Medical School, United States of America

Breast Cancer care includes prevention, early detection, diagnostics and therapeutics. Therapeutic decisions are made based on traditional prognostic factors including tumor size, lymph node status, and factors obtained from pathological assessment including tumor grade, immunohistochemical profile of Estrogen and Progesterone Receptor (ER and PR) and Her2/neu gene amplification status. Point of care technology is not currently used in this routine pathological assessment, but there are new opportunities to expedite and facilitate diagnosis, primarily driven by the need to provide breast cancer diagnoses in low-resource settings to the tens of thousands of women who develop breast cancer worldwide. This presentation will discuss alternative methods of tissue biopsy, handling, and imaging and prognostic marker evaluation that can obviate the need for expensive traditional processing equipment and microscopes, and can allow for more rapid cancer diagnosis and biomarker evaluation compared with current traditional methods.

Paper-based Analytical Devices For the Diagnosis and Monitoring of Infectious Diseases at the Point of Care
Jean-Manuel Segura, Professor, University of Applied Sciences and Arts Western Switzerland Valais, Switzerland

The diagnosis and treatment of infectious diseases require many in vitro diagnostic (IVD) tests to guarantee an accurate and efficient intervention. First, testing of generic inflammation markers such as CRP enable a rapid determination of the origin of the infection, either bacterial or viral. Then detailed analysis of genetic material or of specific biomarkers enables identification of the pathogen in order to select the most appropriate treatment. Finally, monitoring of the administrated therapeutic drugs in the blood allows for personalizing the dose in order to optimize efficacy while avoiding side effects. Performing these different tests at the point of care provides many benefits, in particular a more rapid intervention.

Paper offers many advantages as a material to produce IVD tests as it is cheap, amenable for mass manufacturing, easy to dispose after performing the test and it provides many functionalities for designing a test such as capillary-driven motion of liquids. Two main types of paper-based analytical tests exist with the lateral-flow assays, ideal to perform immunoassays, and the microfluidic paper-analytical devices, microPAD, ideal to perform enzymatic assays.

I will present our most recent results in developing various paper-based IVD tests for the diagnosis and treatment of infectious diseases at the point of care. A particular emphasis will be put on a microPAD test for the monitoring of therapeutic antibiotics in the blood.

Low-Q-Whispering Gallery Modes-A New Platform For Label-Free Investigation of Surface Processes
Lars Dähne, Founder & Managing Director, Surflay Nanotec GmbH, Germany

Low Q whispering gallery modes (WGM) are light waves that circulate up to 15 000 times inside spherical microparticles by total reflection on the inner surface, thereby sensitively scanning the surface. When molecules are adsorbed on the surface of the particles, there is a change in refractive index, leading to shifts of resonance frequency of the WGM. The shift is readout by a high-resolution spectrometer and can be used to quantify surface interactions and their kinetics. In contrast to other WGM setups in toroids or High Q WGM in larger silica particles where the light is coupled in by tangentially arranged light guides, we are using fluorescent molecules inside of 10 µm polymer particles for the generation of the WGM. Consequently the sensor can be activated and readout contactless in smallest cavities like in three dimensional cell arrays or even inside of living cells.

For the specific detection of biomolecules, the WGM sensors are functionalized with antigens or oligonucleotides. An arbitrary number of particles are immobilized inside a chip for which several possibilities are available. After incubation with the analyte, for example a serum sample, the adsorption of complementary molecules is readout as a shift of the WGM signals without the need of special washing or subsequent incubation with “secondary” antibodies. One measurements point require less than 0.1 second per particle. Using large numbers of sensor particles enables high statistical relevance, as well as parallel analysis of almost unlimited number of complementary biomolecules. The talk presents the Low Q WGM technology and indicates its advantages by some application examples.

Networking Reception with Beer and Wine in the Exhibit Hall -- Meet Colleagues and Network with Exhibitors

Close of Day 1 of the Conference

Morning Coffee and Tea Plus Networking in the Exhibit Hall

Monitoring and Diagnosis of Bacterial Infections, Using Micro and Nanotechnology
Winnie Svendsen, Professor, Technical University of Denmark, Denmark

Planar Waveguide Biosensor Technology for Next Generation Point of Care Diagnostics
Reuven Duer, Founder & CEO, Proactive Diagnostics, Inc., United States of America

I’ll be presenting a novel and disruptive detection technology for Point of Care diagnostics.  The presentation includes the underlying physics, implementation solutions and testing data of this unique technology platform.

Smart Thermometers: Screening for Biomarkers Using Molecularly Imprinted Polymers (MIPs) Combined with Thermal Detection
Marloes Peeters, Deputy Director of Chemical Engineering, Newcastle University, United Kingdom

Molecularly Imprinted Polymers (MIPs) are synthetic antibody mimics: similar to antibodies they possess high affinity for a chosen template molecule. However, they have distinct advantages over their natural counterparts such as low-cost, superior chemical and thermal stability, and straightforward production process. In this contribution, we will discuss the use of nanoMIPs that are synthesized via a solid-phase approach. The high affinity nanoparticles prepared by this technique are water-soluble, meaning it is simple to directly functionalize them onto thermocouples via dipcoating. These functionalized thermocouples were subsequently inserted into a home-made heat-transfer device that measures the temperature of the liquid in a flow cell. It was shown that binding of the target to the MIP layer increased the resistance at the solid-liquid interface, leading to a lower temperature being recorded by the functionalized thermocouple. With this method, it was possible to measure various compounds, ranging from small targets such as the antibiotic vancomycin to larger biomarkers such as troponin, in the low nanomolar regime in buffered solutions. The sensitivity was significantly enhanced by using standard EDC coupling to covalently attach the nanoparticles to the surface, allowing to control the surface architecture. Finally, the design of the current flow cell was adjusted to incorporate four thermocouples, demonstrating the first multiplex format of this sensor platform. In this presentation, a case study on the detection of cardiac biomarkers, including troponin, will be discussed.

Embracing Chaos – A Simplified Platform for Multiplexing Digital Assays in Polydisperse Droplets
Samantha Byrnes, Research Scientist, Intellectual Ventures Laboratory, United States of America

We describe a digital assay platform that uses a novel statistical framework applied to assays performed in polydisperse droplets.  We have developed methods for digital culture, PCR, and immunoassays as well as a new technique for increasing multiplexing capabilities.

Morning Coffee and Tea Break and Networking in the Exhibit Hall

Manufacturable Microfluidic Devices – Fine Tuning the Biochemistry and Fabrication
Rosanne Guijt, Professor, Deakin University, Australia

Manufacturability is one of the bottlenecks in the development of commercially viable microfluidic systems. As part of the development of a sample in/answer out assay for the detection of Avian influenza (H1N1) as example, different strategies were used to aid manufacturability. To simplify hardware and minimize thermal stress, isothermal amplification strategies were preferred, and Nucleic Acid Sequence Based Amplification (NASBA) was selected as amplification process because of its high fidelity in replicating RNA. Despite the isothermal amplification at 42C, this is preceded by a thermal annealing step >60C. Chemical and biochemical approaches of circumventing this thermal denaturation step will be compared. The simplified assay uses an external heater, moving magnet for RNA transport and LED induced fluorescence detection, all accessing eh amplification chamber. To minimize geometrical constraint, a dedicated mould was designed for injection moulding of a smooth and straight surface, facilitating side illumination. The described assay facilitated the detection of H1N1 down to ct28.

Ultrafast and Sensitive qPCR-based Detection of Group B Streptococcus (GBS) in Patient Samples in a Matter of Minutes
Maarten Fauvart, R&D Team Leader / Senior Scientist Life Science Technologies, IMEC, Belgium

We present a mass-producible silicon chip-based system for qPCR detection of group B streptococci in five minutes. The analytical performance of the ultrafast qPCR system matches that of the benchtop assay based on a comparison using over 100 patient samples.

The Legal and Regulatory Landscape of Diagnostics Devices
Erik Vollebregt, Partner, Axon Lawyers, Netherlands

The legal and regulatory landscape of diagnostics devices in the EU is in full flux as a result of the In Vitro Diagnostics Regulation, the General Data Protection regulation and other developments like the Brexit. This presentation will address where the rules are at this moment and what this means for the diagnostics industry.

Revisiting the Point of Care – Changes to U.S. FDA Policies on CLIA Waivers and Opportunities in POCT
James Boiani, Partner, Epstein Becker & Green, P.C., United States of America

There have recently been substantial changes for the better in U.S. FDA’s approach to application of CLIA Waiver standards, reflected in new FDA guidance.  Mr. Boiani – a seasoned life sciences attorney and the Founder and General Counsel of the Coalition for CLIA Waiver Reform – will offer his perspectives on FDA’s new approach, the history of its evolution and legal underpinnings, and the opportunities it creates for innovation in the U.S. market.  He will cover key elements of new FDA guidance, including use of agreement studies, human factors evaluations, and dual marketing and CLIA waiver submissions, which could make for a more efficient and accepting regulatory environment.

Networking Lunch in the Exhibit Hall and Poster Viewing

Luncheon Technology Spotlight Presentation: BioDot, Inc. Provides Non-contact, Low Volume Dispensing Solutions Valuable in the Biosensor Industry
Chris Fronczek, Director of Applications, BIODOT, Inc.

Low volume dispensing and liquid handling techniques.

Afternoon Coffee and Tea Break and Networking in the Exhibit Hall

Poster Awards

Novel Bioinstrumentation for Organ to Single Cell Imaging
Barbara Smith, Assistant Professor, School of Biological and Health Systems Engineering, Arizona State University, United States of America

Translation of medical imaging technologies has gained considerable interest over the past decades due, in great part, to the wide range of potential clinical and research applications. Failure of current technologies to provide real-time information does not stem from either a lack of medical interest or importance; rather, it is sourced in four major difficulties: i) miniaturization, ii) automation, iii) speed of acquisition, and iv) integration into commercial tools. This presentation will introduce our recent work investigating novel photoacoustic and ultrasound technologies for use in a variety of unique biomedical applications. The overarching goal is to translate technologies developed in the lab for improved patient outcomes and fundamental knowledge gained.

Enzymatic Biosensor to Support the Diagnosis of Adrenal Gland Tumors
Michael Schöning, Professor and Director, Institute of Nano- and Biotechnologies, Aachen University of Applied Sciences, Germany

Primary aldosteronism (PA) is a disorder in which aldosterone production is inappropriately high and causes hypertension. The clinical guidelines advocate using adrenal vein sampling (AVS) to diagnose PA. However, since AVS is a demanding procedure in which correct cannulation of adrenal veins is challenging, it is used only in a few centers worldwide. To facilitate this medical examination, the detection of adrenaline during AVS could be used as an indicator for the correct position of the catheter, since the adrenaline concentration in adrenal blood (100 nM) is about 100 times higher in comparison to peripheral blood (1 nM).

Two different biosensor approaches for the detection of adrenaline will be presented, which are based on the substrate recycling principle. The first variant is based on an amperometric bi-enzyme biosensor with a genetically modified laccase and a glucose dehydrogenase (GDH). The second example represents a chip-based biosensor with the enzyme PQQ-GDH. Both sensor types have been optimized under laboratory conditions and successfully applied for first measurements in real blood samples, to define an appropriate catheter position.

The SnapChip™ for a Precise Chip-to-Chip Transfer of Nanodroplets
Sebastien Bergeron, Founder and CEO, Parallex BioAssays Inc., Canada

Multiplexed sandwich immunoassays are a powerful technique to measure multiple protein concentrations simultaneously. However, multiplexed assays are prone to cross reactivity effects and can generate false positive signals due to a lack of antibody specificity and selectivity. This is a known weakness of multiplexed assays that is mitigated by months of assay optimization and validation.

Parallex BioAssays replaced the “detection soup” of antibodies by an array of nanodroplets and offers a new CHIP-to-CHIP approach to deliver the detection antibodies precisely to their cognate spots of capture antibodies. As a result, a cross-reaction free chip consisting of an assembly of parallelized assays, physically isolated from each other. The advantages include the possibility to mix-and-match assays and create unique combinations, such as isoforms and phospho-specific antibodies side-by-side. This makes our approach to cell signaling studies (e.g. Akt activation pathway) a rapid and easy alternative to performing dozens of western blots.

The SnapChip™ represents a new format of liquid handling platforms. Beyond exclusive immunoassays, it creates the opportunity to miniaturize and multiplex enzyme activity and cell-based assays in nano-compartments. Current and future applications will be discussed.

Translating Microfluidic Devices from the Academic Benchtop to the Point of Care via Extrusion-Based 3D Printing
E. Brandon Strong, NSF Graduate Research Fellow, California Polytechnic State University, San Luis Obispo, United States of America

While there has been widespread research on microfluidic devices for a wide array of applications, including point-of-care diagnostics, there has been limited translation of these technologies to real-world settings. A significant reason is that current microfabrication techniques (i.e., soft lithography) typically require complex, labor-intensive processes, and necessitate the use of highly-trained personnel. The purpose of our investigation was to fabricate complex microfluidics structures via extrusion-based 3D printing with commercially-available materials ($15-100/kg) & equipment ($200 printer), thereby increasing accessibility to a wider range of researchers. Functional open-faced microfluidics channels less than 100 µm in width (200 µm depth) were readily reproduced, while channels as small as 15 µm in width were also fabricated. All channels were tested without the use of an external pump. Fully enclosed horizontal (200-500 µm) and vertical (500-1000 µm) channels were also fabricated. Hybrid devices contained both vertical and horizontal channels to create complex 3D fluidic arrays. Multi-layered electronic devices with multi-electrode microfluidic wells were created and allowed for simple electroanalysis. 3D printed electronic circuits were also used to thermally actuate paraffin valves in microfluidics channels, which allowed for increased fluid control. Applications in point-of-care diagnostics will be discussed.

Close of Day 2 of the Conference