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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
61

Entwicklung integrierter mikrofluidischer Aktoren für den Einsatz in bioanalytischen Systemen

Nestler, Jörg 21 December 2010 (has links)
In der vorliegenden Arbeit wird eine integrierbare Pumpentechnologie für polymerbasierte mikrofluidische Systeme entwickelt. Ausgehend von den Anforderungen für die Durchführung molekulardiagnostischer Nachweise kommen dabei Fertigungsverfahren zum Einsatz, die sich auch für Einweg-Anwendungen eignen. Das genutzte Aktorprinzip für die integrierten Mikropumpen basiert auf der Elektrolyse von Wasser. Zur besseren technologischen Integrierbarkeit wird das Wasser in Form eines Hydrogels appliziert. Der Elektrolyt wird dabei mit einer Polymermembran mit geringer Wasserdampfdurchlässigkeit verschlossen. Die Membran wird in ihrem plastischen Verformbereich genutzt. Zur Dimensionierung der Mikropumpen und des mikrofluidischen Systems werden analytische und numerische Modelle entwickelt, die eine gute Übereinstimmung mit den Messwerten zeigen. Die Funktionsfähigkeit wird anhand zweier vollständig integriert ablaufender Immunoassays demonstriert. Dabei kommt ein polymerbasierter, optischer Biosensor zum Einsatz.
62

Tetra-Responsive Grafted Hydrogels for Flow Control in Microfluidics

Gräfe, David 25 January 2017 (has links)
Microfluidics covers the science of manipulating small quantities of fluids using microscale devices with great potential in analysis, multiplexing, automation and high-throughput screening. Compared to conventional systems, microfluidics benefits from miniaturization resulting in shortened time of experiments, decreased sample and reagent consumptions as well as reduced overall costs. For microfluidic devices where further weight and cost reduction is additionally required, stimuli-responsive hydrogels are particularly interesting materials since they can convert an environmental stimulus directly to mechanical work without any extra power source. Hydrogels are used as chemostats, micropumps, and chemo-mechanical valves in microfluidics. Existing studies about hydrogels for flow control reported on hydrogels responsive to only one stimulus, including temperature, pH value, and solvent. Combining temperature and pH stimuli within one material is an interesting approach, which allows internal as well as external flow control and broadens potential applications. Among the variety of temperature- and pH-responsive monomers, N-isopropylacrylamide (NiPAAm) and acrylic acid (AA) are considered as ideal building blocks to obtain a hydrogel with pronounced stimuli response. There are different architectures for realizing a temperature- and pH-responsive hydrogel with NiPAAm and AA (e.g. copolymer gels, interpenetrating polymer networks (IPNs), semi-IPNs, or graft copolymer gels). Each approach has its inherent benefits and disadvantages. Grafted hydrogels with a temperature-responsive backbone and pH-responsive graft chains are a promising architecture overcoming drawbacks of copolymer gels (loss of thermoresponsive behavior due to the comonomer), interpenetrating polymer networks (IPNs, difficult fabrication of structured particles via soft lithography), and semi-IPNs (leakage of penetrating polymer). However, studies about multi-responsive grafted hydrogels for flow control in microfluidics are comparatively rare and further research is needed to emphasize their real potential. For this reason, the overall aim of this work was the synthesis of temperature- and pH-responsive grafted hydrogels based on NiPAAm and AA for flow control in microfluidics. This required the synthesis of a pH-responsive macromonomer by RAFT polymerization. As a suitable chain transfer agent with a carboxylic acid group for an end-group functionalization, 2-(dodecyl-thiocarbonothioylthio)-2-methylpropionic (DTP) acid was employed. The approach towards the synthesis of the pH-responsive macromonomer based on two key steps: (i) attaching a functional group, which retains during RAFT polymerization, and (ii) conducting the RAFT polymerization to synthesize the pH-responsive macromonomer. In total, four functionalizations for the macromonomer were investigated, including allyl, unconjugated vinyl, acrylamide, and styrene. End-group analysis and solubility tests revealed that macromonomers with a styrene functionalization are suitable for the synthesis of graft copolymer gels. A series of grafted net-PNiPAAm-g-PAA-styrene hydrogels with a PNiPAAm backbone and PAA-styrene graft chains (Mn = 4200 g/mol, Mw/Mn = 1.6) were prepared and characterized. The main goal was to identify suitable stimuli for an application as a chemo-mechanical valve and to show reversibility of the swelling and shrinking process. Importantly, the temperature sensitivity should be retained, while a pH response needs to be introduced. Equilibrium swelling studies quantified with the response ratio revealed that a grafting density of PAA-styrene between 0.25 and 1 mol-% provides a suitable response towards temperature, pH, salt, and solvent. Furthermore, the swelling and shrinking process is highly reproducible over four consecutive cycles for all four stimuli. In order to evaluate the swelling kinetics of grafted net-PNiPAAm-g-PAA-styrene hydrogels, the collective diffusion model extended by a volume specific surface was applied. The determined cooperative diffusion coefficients of net-PNiPAAm-g-PAA-styrene indicated faster response time with increasing PAA-styrene content. Remarkably, net-PNiPAAm-g-PAA-styrene containing 1 mol-% PAA-styrene exhibited an accelerated swelling rate by a factor of 9 compared to pure net-PNiPAAm. Rheological analysis of net-PNiPAAm-g-PAA-styrene showed that an increasing graft density leads to decreasing mechanical stability. The photopolymerization experiments showed that the gelation time linearly increases with the grafting density. Grafted net-PNiPAAm-g-PAA-styrene hydrogels were tested in two fluidic setups for flow control. A straightforward fluidic platform was developed consisting of a fluid reservoir, an inlet channel, an actuator chamber and an outlet channel. The actuator chamber was filled with crushed hydrogel particles. Accordingly, the fluid flow was directed by the active resistance of the hydrogel particles in the actuator chamber (i.e. swelling degree) and allowed flow control by the local environmental conditions. Flow rate studies showed that the fluid flow throttles when the inlet channel was provided with a solution in which the hydrogel swells (pH 9 buffer solution at room temperature). In contrast, the hydrogel-based valve opens immediately when a solution was used in which the hydrogel collapses. The advantageous properties of net-PNiPAAm-g-PAA-styrene were highlighted by using pH, salt and solvent stimulus in one experiment. Remarkably, the opening and closing function was reversible over six consecutive cycles. As part of a collaboration project with the chair of polymeric microsystems within the Cluster of Excellence Center for Advancing Electronics Dresden (A. Richter and P. Frank), membrane assures hydraulic coupling in a chemo-fluidic membrane transistor (CFMT) and grafted net-PNiPAAm-g-PAA-styrene hydrogels were combined to emphasize the potential of both systems. Flow rate studies showed that 4 different stimuli can be used to control the opening and closing state of the CFMT. Multiple opening and closing cycles revealed no considerable changes in the valve function emphasizing a high potential for an application in microfluidics.
63

Multifunktionsfeldeffekttransistoren zur Strömungs-, Chemo- und Biosensorik in Lab on a Chip-Systemen

Truman Sutanto, Pagra 14 December 2007 (has links)
In dieser Arbeit wird eine neue Methode und ein neuartiges FET -Sensorelement zum Nachweis von Flüssigkeitsbewegungen vorgestellt, das zudem bei Bedarf auch als Chemo- oder Biosensor fungieren kann. Das Einsatzspektrum von FET-basierten Sensoren in Lab on a Chip-Systemen wird dadurch entscheidend erweitert. Bei dem entwickelten FET-Sensor Bauelement handelt es sich um einen normally-on n-leitenden Dünnschichtfeldeffekttransistor mit Ti-Au-Kontakten, basierend auf Silicon-on-Insulator- Substraten, wobei das natürliche Oxid des Siliziumfilms als Schnittstelle zum Elektrolyten bzw. zur Flüssigkeit verwendet wird. Der mit 10exp16 Bor Atomen pro cm³ p-dotierte Siliziumdünnfilm hat eine Dicke von nur 55 nm und ist durch eine 95 nm dicke Siliziumdioxidschicht vom darunterliegenden Siliziumsubstrat von 600 µm Dicke elektrisch isoliert. Aufgrund der geringen Schichtdicke durchdringt die feldempfindliche Raumladungs- bzw. Verarmungszone die gesamte Dünnschicht, so dass durch Anlegen einer Backgatespannung am Substrat der spezifische Widerstand und die Empfindlichkeit des Bauelements eingestellt werden können. Grundlegende ISFET-Funktionalitäten wie die Empfindlichkeit auf Änderungen der Ionenstärke und des pH-Wertes werden nachgewiesen und ein ENFET-Glukosesensor realisiert. Zudem wird im Hinblick auf die Separation von Emulsionen der Nachweis erbracht, dass die Benetzung mit Hexan und Toluol eine Änderung der spezifischen Leitfähigkeit bewirkt, und die Empfindlichkeit des Bauelements nach Beschichtung mit einem hydrophoben Methacrylatcopolymerfilm erhalten bleibt. Hinsichtlich der Verwendung des FET-Sensor Bauelements zum Nachweis von Flüssigkeitsbewegungen wird zunächst ein theoretisches Modell entwickelt, dessen Kernaussage ist, dass sich in einem rechteckigen Kanal der relative Bedeckungsgrad mit Flüssigkeit direkt proportional zum Drainstrom des FET-Sensors verhält. Basierend auf diesem theoretischen Modell, welches experimentell belegt wird, können mittels eines einzelnen FET-Sensors Füllstand und Füllgeschwindigkeit bzw. bei bekannter Füllgeschwindigkeit Kapillarvolumen und Kapillargeometrie bestimmt werden. Abweichungen von der direkten Proportionalität erlauben zudem, Rückschlüsse auf die Benetzungseigenschaften der Kapillaren und die Dynamik an der Halbleitergrenzfläche zu ziehen. Ist ein Sensorelement vollständig mit Flüssigkeit bedeckt, wird mittels Lösungsmitteltropfen als Markerobjekten die Strömungsgeschwindigkeit bestimmt. Ändert sich die Ionenkonzentration im Elektrolyten als Funktion der Strömungsgeschwindigkeit, so kann die Strömungsgeschwindigkeit durch Messung der Ionenkonzentration mittels FET-Sensor ebenfalls ermittelt werden. Als wichtigster Demonstrator für die Verwendung des FET-Sensors wird ein komplexes Lab on a Chip-System zur Separation von Emulsionen auf chemisch strukturierten Oberflächen entwickelt, bei dem der Separationsvorgang mittels FET-Sensorarray verfolgt werden kann. Zur einfachen Herstellung chemisch modifizierter Oberflächen für die Separationsexperimente werden die Abscheidung von nanoskaligen hydrophoben Methacrylatcopolymerfilmen und die selektive Fluorsilanisierung von Oberflächen sowie deren Lösungsmittelbeständigkeit in Wasser, Toluol und Aceton untersucht. Dabei zeigt sich, dass die Hydrophobie nach Lösungsmittelbehandlung weitestgehend erhalten bleibt, Wasserrückstände im Methacrylatfilm aber zu einer reversiblen Schichtdegradation führen können. Als Modellsystem werden Hexan-Wasser- bzw. Toluol-Wasser-Emulsionen verwendet, die auf Oberflächen getrennt werden, deren eine Seite hydrophil, und deren andere Seite hydrophob ist (Stufengradient). Der Separationsprozess beruht auf der großen Affinität des Wassers hin zu polaren Oberflächen, wobei das wenig selektive Lösungsmittel zur unpolaren Seite gedrängt wird. Zur Erlangung eines tieferen Verständnisses des Prozesses werden die Tropfenkoaleszenz und der Einfluss geometrischer Beschränkungen untersucht. Die Versuche werden sowohl auf offenen Oberflächen als auch im Spalt, unter Verwendung von hydrophilen und hydrophoben Oberflächen, durchgeführt. Es zeigt sich, dass sich die Dynamik der Tropfenkoaleszenz im Spalt umgekehrt zur Dynamik auf offenen Oberflächen verhält. Dies wird mittels eines hierzu entwickelten theoretischen Modells erklärt, welches die Minimierung der Oberflächenenergie und Hystereseeffekte einbezieht. Das Lab on a Chip-System schließlich besteht aus einem mit Siliziumnitrid beschichteten FET-Sensorchip, auf den eine Separationszelle aufgeklebt ist. Neben dem Einlass für die Emulsion ist ein weiterer Einlass vorhanden, durch den Salzsäure für eine pH-Reaktion zugegeben werden kann. Der gesamte Separationsprozess sowie die anschließende pH-Reaktion, lassen sich bequem am PC anhand der Änderung der Stromstärke der einzelnen Sensoren verfolgen und analysieren. Wichtige Ergebnisse hier sind: 1) Mittels eines quasi 1-dimensionalen Sensorarrays kann der Verlauf einer Flüssigkeitsfront in einem 2-dimensionalen Areal überwacht bzw. dargestellt werden. 2) Anhand der Signatur des Signalverlaufs bei pH-Änderung und Flüssigkeitsbewegung, können beide Prozesse unterschieden werden. Der Sensor kann also zum Nachweis von Flüssigkeitsbewegungen und zugleich als Chemosensor eingesetzt werden. Es wurde also nicht nur ein neuartiges, äußerst robustes, chemikalienbeständiges und biokompatibles Multifunktionssensorelement mit Abmessungen im Mikrometer- bis Millimeterbereich entwickelt, sondern auch eine neue Methode entwickelt, mit der es möglich ist, sowohl (bio-)chemische Reaktionen als auch die Bewegung von Flüssigkeiten in Lab on a Chip-Systemen nachzuweisen.
64

Mechanical behavior and pore integration density optimization of switchable hydrogel composite membranes

Ehrenhofer, Adrian, Hahn, Manfred, Hofmann, Martin, Wallmersperger, Thomas 19 March 2021 (has links)
Switchable hydrogel-layered composite membranes can be used for the analysis of particle size distributions. This functionality is provided by pores with controllable diameter. In order to obtain a device that can be used to measure the cell size distribution in native biological samples, lots of switchable pores are required. In the current work, we model and simulate the mechanical behavior of active composite membranes with switchable pores. This is done in order to find the maximum number of pores that can be integrated into a membrane without cross-influencing effects on the actuation of the pores. Therefore, we investigate (1) the interaction of active pores inside the multifunctional composite and (2) the membrane bending under microfluidic pressure load. We show that through miniaturization, sufficient pores can be added to a permeation control membrane for processing native blood samples. The envisioned device allows a parallelized measurement of cell sizes in a simple lab-on-a-chip setup.
65

Adjustable fluid and particle permeation through hydrogel composite membranes

Ehrenhofer, Adrian, Wallmersperger, Thomas 24 March 2021 (has links)
Membranes act as smart structures in respect to their permeation abilities. Control of particle and fluid permeation through a synthetic membrane can be achieved by using different effects like size-exclusion or electromagnetic interactions that occur between the particles and membrane pores. The simulation of controlled permeability provides an insight into the smart behavior of membranes for chemical signal processing, sensing interfaces or lab-on-a-chip devices. In the current work, we model the underlying physical processes on a microfluidic level using the engineer’s approach of laminar flow through pipes. Different pore geometries inside a composite membrane system consisting of a polyethylene terephthalate support membrane and a poly(N-isopropylacrylamide) hydrogel-layer are investigated. Simulations for different states of thermally induced pore opening are performed for free and blocked states. From the results we derive paradigms for the design of a membrane system for microfluidic cell-size profiling considering stimulus-range, pore shape and measurement setup.
66

Cellulose nanofibril-based Layer-by-Layer system for immuno-capture of circulating tumor cells in microfluidic devices

Lahchaichi, Ekeram January 2021 (has links)
År 2020 listade Världshälsoorganisationen (WHO) cancer som den globalt ledande dödsorsaken med över 10 miljoner dödsfall årligen. Av dessa 10 miljoner fall förekommer nästan 70% i låg- till medelinkomstländer - en siffra som på grund av den låga prioriteringen av cancerbehandling- och diagnostik förväntas öka till 85% redan år 2030. Att utveckla enkla, specifika och prisvärda verktyg för diagnostik kommer därför att bli avgörande för förebyggandet av cancer på en global nivå. För att komma ett steg närmare denna utveckling optimerades och testades i denna studie ett mikrofluidiskt system, utvecklat genom layer-bylayer- metoden, baserat på cellulosa nanofibriller med förmågan att isolera och fånga cirkulerande tumörceller. För att uppnå en termodynamisk jämvikt optimerades systemets hydrodynamiska parametrar optimerades för att uppnå en homogen fördelning med hög densitet av det cellulosa-baserade systemet i det mikrofluidiska chippet. Då jämvikt är grundläggande för att maximera det efterföljande beläggningen av antikroppar, och därmed hur effektivt celler isoleras, modifierades parametrar såsom koncentration, flödeshastighet, inkubationstid med fler tills att önskad effekt uppnåtts. Således koncepttestades systemet genom att fånga celler spetsade i blod och därmed demonstrera att systemet kan användas i syfte att isolera cancerceller från blodprov. Detta öppnar upp för utveckling av liknande diagnostiska verktyg som kan användas för att isolera lågfrekventa celler direkt från blod. / In 2020, the World Health Organization (WHO) listed cancer as the leading cause of death worldwide, reaching a staggering number of 10 million cancer-related deaths annually. Of these 10 million deaths, nearly 70% occurred in low- and middle-income countries; a number that is expected to increase to 85% by 2030 due to the lack of resources as well as low priority of the development of cancer treatment and diagnosis. Hence, the development of a sophisticated, specific and affordable diagnostic tool will be crucial for global cancer prevention and control. In this study, a cellulose nanofibril-based Layer-by-Layer system for immuno-capture of tumour cells in a microfluidic device was optimized and tested for the development of a simple and cost-effective diagnostic tool for use in resource-limited areas. In the pursuit of a thermodynamic equilibrium, the hydrodynamic parameters of the system were optimized to achieve a homogeneous distribution with a high surface density of the cellulose-based system across the microfluidic channels. Since an equilibrated system is essential to maximize the antibody coating, and thereby cell capture efficiency, parameters including but not limited to concentration, flow rate and incubation time were altered until a desired effect had been achieved. Thus, as proof-of-concept, the system was tested by capturing cancer cells spiked into whole blood, thereby demonstrating that the system can be utilized for the purpose of isolating cancer cells from blood samples. This paves the way for the development of similar clinical diagnostic tools for the isolation of rare cells directly from whole blood.
67

Affinity Based Capture of Circulating Tumour Cells Using Designed Ankyrin Repeat Proteins (DARPins) in a Microfluidic System

Spåre, Emil January 2021 (has links)
Designade ankyrinupprepningsproteiner (DARPiner) är små, mycket stabila antikroppsmimetiska proteiner. I det här projektet användes anti-EpCAM-DARPiner tillsammans med mikrofluidik för att avgära om de kunde fånga upp HCT116-celler mer effektivt än anti-EpCAM-antikroppar. Ytorna på insidan av mikroffluidikkanaler förändrades genom bindning av N-γ-maleimidobutyryl-oxysuccinimidester (GMBS) och merkaptopropyltrietoxysilan (MPTES) för anti-EpCAM-antikroppar och GMBS och (3-aminopropyl)trietoxysilan (APTES) för DARPiner. Båda kanaltyperna testades genom inflöde av cancerceller och helblod blandat med cancerceller. Ingen effektiv och konsekvent celluppfångst åstadkoms trots att det visades att antikropparna och DARPinerna kunde binda till cellerna direkt och att test med fluorescenta DARPiner och antikroppar visade att ytförändringskemin var fungerande. Slutsatsen blev att de mest troliga orsakerna till misslyckandena var att ytförändringskemin påverkade proteinernas bindningsförmåga negativt eller att proteinerna bands till kanalernas yta i fel riktning. DARPiner är fortfarande intressanta för tillämpningar inom mikrofluidik, men vidare förbättring av det experimentella protokollet behövs. / Designed ankyrin repeat proteins (DARPins) are small and highly stable antibody mimetics. In this project, anti-EpCAM DARPins were used in conjunction with microfluidics to determine if they could capture HCT116 cells more effectively than anti-EpCAM antibodies. The inside surfaces of microfluidic chips were modified using N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS) and mercaptopropyltriethoxysilane (MPTES) for anti-EpCAM antibodies, and surface modifications for anti-EpCAM DARPins were made using GMBS and (3-aminopropyl)triethoxysilane (APTES). Both chip types were tested using cancer cells and whole blood mixed with cancer cells. No effective and consistent cell capture was achieved, despite the antibodies and DARPins being shown to be able to bind to the cells directly and tests with fluorescently labelled DARPins and antibodies showing that the surface modification chemistry used was functional. It was concluded that the most likely causes of the failures were surface modifications interfering with the binding ability of the proteins, or improper orientation of the bound proteins. The DARPin remains a protein of interest for microfluidic applications, but further changes and optimisation of the experimental protocol is necessary.
68

Mechanical behavior and pore integration density optimization of switchable hydrogel composite membranes

Ehrenhofer, Adrian, Hahn, Manfred, Hofmann, Martin, Wallmersperger, Thomas 11 August 2020 (has links)
Switchable hydrogel-layered composite membranes can be used for the analysis of particle size distributions. This functionality is provided by pores with controllable diameter. In order to obtain a device that can be used to measure the cell size distribution in native biological samples, lots of switchable pores are required. In the current work, we model and simulate the mechanical behavior of active composite membranes with switchable pores. This is done in order to find the maximum number of pores that can be integrated into a membrane without cross-influencing effects on the actuation of the pores. Therefore, we investigate (1) the interaction of active pores inside the multifunctional composite and (2) the membrane bending under microfluidic pressure load. We show that through miniaturization, sufficient pores can be added to a permeation control membrane for processing native blood samples. The envisioned device allows a parallelized measurement of cell sizes in a simple lab-on-a-chip setup.
69

Simulation of controllable permeation in PNIPAAm coated membranes

Ehrenhofer, Adrian, Wallmersperger, Thomas, Richter, Andreas 06 August 2019 (has links)
Membranes separate uid compartments and can comprise transport structures for selective permeation. In biology, channel proteins are specialized in their atomic structure to allow transport of specific compounds (selectivity). Conformational changes in protein structure allow the control of the permeation abilities by outer stimuli (gating). In polymeric membranes, the selectivity is due to electrostatic or size-exclusion. It can thus be controlled by size variation or electric charges. Controllable permeation can be useful to determine particle-size distributions in continuous ow, e.g. in micro uidics and biomedicine to gain cell diameter profiles in blood. The present approach uses patterned polyethylene terephthalate (PET) membranes with hydrogel surface coating for permeation control by size-exclusion. The thermosensitive hydrogel poly(N-isopropylacrylamide) (PNIPAAm) is structured with a cross-shaped pore geometry. A change in the temperature of the water ow through the membrane leads to a pore shape variation. The temperature dependent behavior of PNIPAAm can be numerically modeled with a temperature expansion model, where the swelling and deswelling is depicted by temperature dependent expansion coefficients. In the present study, the free swelling behavior was implemented to the Finite Element tool ABAQUS for the complex composite structure of the permeation control membrane. Experimental values of the geometry characteristics were derived from microscopy images with the tool ImageJ and compared to simulation results. Numerical simulations using the derived thermomechanical model for different pore geometries (circular, rectangle, cross and triangle) were performed. With this study, we show that the temperature expansion model with values from the free swelling behavior can be used to adequately predict the deformation behavior of the complex membrane system. The predictions can be used to optimize the behavior of the membrane pores and the overall performance of the smart membrane.
70

Lab-on-a-chip platform for high throughput drug discovery with DNAencoded chemical libraries

Grünzner, S., Reddavide, F. V., Steinfelder, C., Cui, M., Busek, M., Klotzbach, U., Zhang, Y., Sonntag, F. 09 August 2019 (has links)
The fast development of DNA-encoded chemical libraries (DECL) in the past 10 years has received great attention from pharmaceutical industries. It applies the selection approach for small molecular drug discovery. Because of the limited choices of DNA-compatible chemical reactions, most DNA-encoded chemical libraries have a narrow structural diversity and low synthetic yield. There is also a poor correlation between the ranking of compounds resulted from analyzing the sequencing data and the affinity measured through biochemical assays. By combining DECL with dynamical chemical library, the resulting DNA-encoded dynamic library (EDCCL) explores the thermodynamic equilibrium of reversible reactions as well as the advantages of DNA encoded compounds for manipulation/detection, thus leads to enhanced signal-to-noise ratio of the selection process and higher library quality. However, the library dynamics are caused by the weak interactions between the DNA strands, which also result in relatively low affinity of the bidentate interaction, as compared to a stable DNA duplex. To take advantage of both stably assembled dual-pharmacophore libraries and EDCCLs, we extended the concept of EDCCLs to heat-induced EDCCLs (hi-EDCCLs), in which the heat-induced recombination process of stable DNA duplexes and affinity capture are carried out separately. To replace the extremely laborious and repetitive manual process, a fully automated device will facilitate the use of DECL in drug discovery. Herein we describe a novel lab-on-a-chip platform for high throughput drug discovery with hi-EDCCL. A microfluidic system with integrated actuation was designed which is able to provide a continuous sample circulation by reducing the volume to a minimum. It consists of a cooled and a heated chamber for constant circulation. The system is capable to generate stable temperatures above 75 °C in the heated chamber to melt the double strands of the DNA and less than 15 °C in the cooled chamber, to reanneal the shuffled library. In the binding chamber (the cooled chamber) specific retaining structures are integrated. These hold back beads functionalized with the target protein, while the chamber is continuously flushed with library molecules. Afterwards the whole system can be flushed with buffer to wash out unspecific bound molecules. Finally the protein-loaded beads with attached molecules can be eluted for further investigation

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