This review presents an overview of 20 years of worldwide development in the field of biosensors based on special types of surface acoustic wave (SAW) devices that permit the highly sensitive detection of biorelevant molecules in liquid media (such as water or aqueous buffer solutions). 1987 saw the first approaches, which used either horizontally polarized shear waves (HPSW) in a delay line configuration on lithium tantalate (LiTaO(3)) substrates or SAW resonator structures on quartz or LiTaO(3) with periodic mass gratings. The latter are termed "surface transverse waves" (STW), and they have comparatively low attenuation values when operated in liquids. Later Love wave devices were developed, which used a film resonance effect to significantly reduce attenuation. All of these sensor approaches were accompanied by the development of appropriate sensing films. First attempts used simple layers of adsorbed antibodies. Later approaches used various types of covalently bound layers, for example those utilizing intermediate hydrogel layers. Recent approaches involve SAW biosensor devices inserted into compact systems with integrated fluidics for sample handling. To achieve this, the SAW biosensors can be embedded into micromachined polymer housings. Combining these two features will extend the system to create versatile biosensor arrays for generic lab use or for diagnostic purposes.
The field of microfluidics or lab-on-a-chip technology aims to improve and extend the possibilities of bioassays, cell biology and biomedical research based on the idea of miniaturization. Microfluidic systems allow more accurate modelling of physiological situations for both fundamental research and drug development, and enable systematic high-volume testing for various aspects of drug discovery. Microfluidic systems are in development that not only model biological environments but also physically mimic biological tissues and organs; such 'organs on a chip' could have an important role in expediting early stages of drug discovery and help reduce reliance on animal testing. This Review highlights the latest lab-on-a-chip technologies for drug discovery and discusses the potential for future developments in this field.
Thermochemical decomposition of organic materials under heat-treatment in the absence of oxygen, known as the pyrolysis process, is often employed to convert micro and nano patterned polymers into carbon structures, which are subsequently used as device components. Pyrolysis is performed at ≥900°C, which entails substrate materials with a high thermal stability that excludes flexible, polymeric substrates. We use optimized laser radiation to pattern graphitic carbon structures onto commercially available polyimide (Kapton) sheets in the micrometer to millimeter scale by inducing a localized, rapid pyrolysis, for the fabrication of flexible devices. Resulting laser carbon films are electrically conductive and exhibit a high-surface area with a hierarchical porosity distribution along their cross-section. The material is obtained using various combinations of laser parameters and pyrolysis environment (oxygen-containing and inert). Extensive characterization of laser carbon is performed to understand the correlation between the material properties and laser parameters, primarily fluence and power. A photothermal carbonization mechanism based on the plume formation is proposed. Further, laser carbon is used for the fabrication of enzymatic, pH-based urea sensors using two approaches: (i) direct urease enzyme immobilization onto carbon and (ii) electrodeposition of an intermediate chitosan layer prior to urease immobilization. This flexible sensor is tested for quantitative urea detection down to 10 −4 M concentrations, while a qualitative, color-indicative test is performed on a folded sensor placed inside a tube to demonstrate its compatibility with catheters. Laser carbon is suitable for a variety of other flexible electronics and sensors, can be conveniently integrated with an external circuitry, heating elements, and with other microfabrication techniques such as fluidic platforms.
The oxyfunctionalization of unactivated C−H bonds can selectively and efficiently be catalyzed by oxygenase-containing whole-cell biocatalysts. Recombinant Escherichia coli W3110 containing the alkane monooxygenase AlkBGT and the outer membrane protein AlkL from Pseudomonas putida GPo1 have been shown to efficiently catalyze the terminal oxyfunctionalization of renewable fatty acid methyl esters yielding bifunctional products of interest for polymer synthesis. In this study, AlkBGTL-containing E. coli W3110 is shown to catalyze the multistep conversion of dodecanoic acid methyl ester (DAME) via terminal alcohol and aldehyde to the acid, exhibiting Michaelis-Menten-type kinetics for each reaction step. In two-liquid phase biotransformations, the product formation pattern was found to be controlled by DAME availability. Supplying DAME as bulk organic phase led to accumulation of the terminal alcohol as the predominant product. Limiting DAME availability via application of bis(2-ethylhexyl)phthalate (BEHP) as organic carrier solvent enabled almost exclusive acid accumulation. Furthermore, utilization of BEHP enhanced catalyst stability by reducing toxic effects of substrate and products. A further shift towards the overoxidized products was achieved by co-expression of the gene encoding the alcohol dehydrogenase AlkJ, which was shown to catalyze efficient and irreversible alcohol to aldehyde oxidation in vivo. With DAME as organic phase, the aldehyde accumulated as main product using resting cells containing AlkBGT, AlkL, as well as AlkJ. This study highlights the versatility of whole-cell biocatalysis for synthesis of industrially relevant bifunctional building blocks and demonstrates how integrated reaction and catalyst engineering can be implemented to control product formation patterns in biocatalytic multistep reactions.
Since the first biosensor was introduced in 1962 by Clark and Lyons, there has been increasing demand for such analytical devices in diagnostic applications. Research initially focussed mainly on detector principles and recognition elements, whereas the packaging of the biosensors and the microfluidic integration has been discussed only more recently. However, to obtain a user-friendly and well-performing analytical device, those components have to be considered all together. This review outlines the requirements and the solutions suggested for the integration of suitable biosensors in packaging and the integration of those encapsulated biosensors into a microfluidic surrounding resulting in a complete and efficient analytical device for diagnostic applications. The components required for a complete biosensor instrument are described and the latest developments which meet the requirements for diagnostic applications, such as single-use components and arrays for multiparameter detection, are discussed. The current state and the future of biosensors in the field of clinical diagnostics are outlined, particularly on the basis of label-free assay formats and the detection of prominent biomarkers for cancer and autoimmune disorders.
Bulk acoustic wave (BAW) and surface acoustic wave (SAW) sensor devices have successfully been used in a wide variety of gas sensing, liquid sensing, and biosensing applications. Devices include BAW sensors using thickness shear modes and SAW sensors using Rayleigh waves or horizontally polarized shear waves (HPSWs). Analyte specificity and selectivity of the sensors are determined by the sensor coatings. If a group of analytes is to be detected or if only selective coatings (i.e., coatings responding to more than one analyte) are available, the use of multi-sensor arrays is advantageous, as the evaluation of the resulting signal patterns allows qualitative and quantitative characterization of the sample. Virtual sensor arrays utilize only one sensor but combine it with enhanced signal evaluation methods or preceding sample separation, which results in similar results as obtained with multi-sensor arrays. Both array types have shown to be promising with regard to system integration and low costs. This review discusses principles and design considerations for acoustic multi-sensor and virtual sensor arrays and outlines the use of these arrays in multi-analyte detection applications, focusing mainly on developments of the past decade.
Biosensors combine a transducer with a biorecognition element and thus are able to transform a biochemical event on the transducer surface directly into a measurable signal. By this they have the potential to provide rapid, real-time, and accurate results in a comparatively easy way, which makes them promising analytical devices. Since the first biosensor was introduced in 1962 as an "enzyme electrode" for monitoring glucose in blood, medical applications have been the main driving force for further biosensor development. In this chapter we outline potential biosensor setups, focusing on transduction principles, biorecognition layers, and biosensor test formats, with regard to potential applications. A summary of relevant aspects concerning biosensor integration in efficient analytical setups is included. We describe the latest applications of biosensors in diagnostic applications focusing on detection of molecular biomarkers in real samples. An overview of the current state and future trends of biosensors in this field is given.
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