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A biosensor is an analytical device, used for the detection of a chemical substance, that combines a biological component with a physicochemical detector. The sensitive biological element, e.g. tissue, microorganisms, , , , antibody, , etc., is a biologically derived material or biomimetic component that interacts with, binds with, or recognizes the analyte under study. The biologically sensitive elements can also be created by biological engineering. The Biotransducer or the detector element, which transforms one signal into another one, works in a physicochemical way: optical, piezoelectric, electrochemical, electrochemiluminescence etc., resulting from the interaction of the analyte with the biological element, to easily measure and quantify. The biosensor reader device connects with the associated electronics or signal processors that are primarily responsible for the display of the results in a user-friendly way. This sometimes accounts for the most expensive part of the sensor device, however it is possible to generate a user friendly display that includes transducer and sensitive element (holographic sensor). The readers are usually custom-designed and manufactured to suit the different working principles of biosensors.
Antibody-antigen interactions can also be used for Serology, or the detection of circulating antibodies in response to a specific disease. Importantly, serology tests have become an important part of the global response to the COVID-19 pandemic.
There also exist some disadvantages of tissues, like the lack of specificity due to the interference of other enzymes and longer response time due to the transport barrier.
Alternatively, three-dimensional lattices (hydrogel/xerogel) can be used to chemically or physically entrap these (whereby chemically entrapped it is meant that the biological element is kept in place by a strong bond, while physically they are kept in place being unable to pass through the pores of the gel matrix). The most commonly used hydrogel is sol-gel, glassy silica generated by polymerization of silicate monomers (added as tetra alkyl orthosilicates, such as TMOS or TEOS) in the presence of the biological elements (along with other stabilizing polymers, such as PEG) in the case of physical entrapment.
Another group of hydrogels, which set under conditions suitable for cells or protein, are Acrylate polymer hydrogel, which polymerizes upon radical initiation. One type of radical initiator is a peroxide radical, typically generated by combining a persulfate with TEMED (Polyacrylamide gel are also commonly used for protein electrophoresis), alternatively light can be used in combination with a photoinitiator, such as DMPA (2,2-dimethoxy-2-phenylacetophenone). Smart materials that mimic the biological components of a sensor can also be classified as biosensors using only the active or catalytic site or analogous configurations of a biomolecule.
Another example, the potentiometric biosensor, (potential produced at zero current) gives a logarithmic response with a high dynamic range. Such biosensors are often made by screen printing the electrode patterns on a plastic substrate, coated with a conducting polymer and then some protein (enzyme or antibody) is attached. They have only two electrodes and are extremely sensitive and robust. They enable the detection of analytes at levels previously only achievable by HPLC and LC/MS and without rigorous sample preparation. All biosensors usually involve minimal sample preparation as the biological sensing component is highly selective for the analyte concerned. The signal is produced by electrochemical and physical changes in the conducting polymer layer due to changes occurring at the surface of the sensor. Such changes can be attributed to ionic strength, pH, hydration and redox reactions, the latter due to the enzyme label turning over a substrate. Field effect transistors, in which the metal gate region has been modified with an enzyme or antibody, can also detect very low concentrations of various analytes as the binding of the analyte to the gate region of the FET cause a change in the drain-source current.
Impedance spectroscopy based biosensor development has been gaining traction nowadays and many such devices / developments are found in the academia and industry. One such device, based on a 4-electrode electrochemical cell, using a nanoporous alumina membrane, has been shown to detect low concentrations of human alpha thrombin in presence of high background of serum albumin. Also interdigitated electrodes have been used for impedance biosensors.
An ion channel switch (ICS) biosensor can be created using gramicidin, a dimeric peptide channel, in a tethered bilayer membrane. One peptide of gramicidin, with attached antibody, is mobile and one is fixed. Breaking the dimer stops the ionic current through the membrane. The magnitude of the change in electrical signal is greatly increased by separating the membrane from the metal surface using a hydrophilic spacer.
Quantitative detection of an extensive class of target species, including proteins, bacteria, drug and toxins has been demonstrated using different membrane and capture configurations. The European research project Greensense develops a biosensor to perform quantitative screening of drug-of-abuse such as THC, morphine, and cocaine in saliva and urine.
Antibodies and artificial families of Antigen Binding Proteins (AgBP) are well suited to provide the recognition module of RF biosensors since they can be directed against any antigen (see the paragraph on bioreceptors). A general approach to integrate a solvatochromic fluorophore in an AgBP when the atomic structure of the complex with its antigen is known, and thus transform it into a RF biosensor, has been described. A residue of the AgBP is identified in the neighborhood of the antigen in their complex. This residue is changed into a cysteine by site-directed mutagenesis. The fluorophore is chemically coupled to the mutant cysteine. When the design is successful, the coupled fluorophore does not prevent the binding of the antigen, this binding shields the fluorophore from the solvent, and it can be detected by a change of fluorescence. This strategy is also valid for antibody fragments.
However, in the absence of specific structural data, other strategies must be applied. Antibodies and artificial families of AgBPs are constituted by a set of hypervariable (or randomized) residue positions, located in a unique sub-region of the protein, and supported by a constant polypeptide scaffold. The residues that form the binding site for a given antigen, are selected among the hypervariable residues. It is possible to transform any AgBP of these families into a RF biosensor, specific of the target antigen, simply by coupling a solvatochromic fluorophore to one of the hypervariable residues that have little or no importance for the interaction with the antigen, after changing this residue into cysteine by mutagenesis. More specifically, the strategy consists in individually changing the residues of the hypervariable positions into cysteine at the genetic level, in chemically coupling a solvatochromic fluorophore with the mutant cysteine, and then in keeping the resulting conjugates that have the highest sensitivity (a parameter that involves both affinity and variation of fluorescence signal). This approach is also valid for families of antibody fragments.
A posteriori studies have shown that the best reagentless fluorescent biosensors are obtained when the fluorophore does not make non-covalent interactions with the surface of the bioreceptor, which would increase the background signal, and when it interacts with a binding pocket at the surface of the target antigen. The RF biosensors that are obtained by the above methods, can function and detect target analytes inside living cells.
Electrochemiluminescence (ECL) is nowadays a leading technique in biosensors. Since the excited species are produced with an electrochemical stimulus rather than with a light excitation source, ECL displays improved signal-to-noise ratio compared to photoluminescence, with minimized effects due to light scattering and luminescence background. In particular, coreactant ECL operating in buffered aqueous solution in the region of positive potentials (oxidative-reduction mechanism) definitively boosted ECL for immunoassay, as confirmed by many research applications and, even more, by the presence of important companies which developed commercial hardware for high throughput immunoassays analysis in a market worth billions of dollars each year.
Thermometric biosensors are rare.
The first BioFET was the ion-sensitive field-effect transistor (ISFET), invented by Piet Bergveld for electrochemical and biological applications in 1970. the adsorption FET (ADFET) was patented by P.F. Cox in 1974, and a hydrogen-sensitive MOSFET was demonstrated by I. Lundstrom, M.S. Shivaraman, C.S. Svenson and L. Lundkvist in 1975. The ISFET is a special type of MOSFET with a gate at a certain distance, and where the metal gate is replaced by an ion-sensitive membrane, electrolyte solution and reference electrode. The ISFET is widely used in biomedical applications, such as the detection of DNA hybridization, biomarker detection from blood, antibody detection, glucose measurement, pH sensing, and genetic technology.
By the mid-1980s, other BioFETs had been developed, including the gas sensor FET (GASFET), pressure sensor FET (PRESSFET), chemical field-effect transistor (ChemFET), ISFET (REFET), enzyme-modified FET (ENFET) and immunologically modified FET (IMFET). By the early 2000s, BioFETs such as the DNA field-effect transistor (DNAFET), gene-modified FET (GenFET) and cell-potential BioFET (CPFET) had been developed.
In biotechnology, analysis of the chemical composition of cultivation broth can be conducted in-line, on-line, at-line and off-line. As outlined by the US Food and Drug Administration (FDA) the sample is not removed from the process stream for in-line sensors, while it is diverted from the manufacturing process for on-line measurements. For at-line sensors the sample may be removed and analyzed in close proximity to the process stream. An example of the latter is the monitoring of lactose in a dairy processing plant.Pasco, Neil; Glithero, Nick. Lactose at-line biosensor 1st viable industrial biosensor? (accessed 30 January 2013). Off-line biosensors compare to bioanalysis that are not operating in the field, but in the laboratory. These techniques are mainly used in agriculture, food technology and biomedicine.
In medical applications biosensors are generally categorized as in vitro and in vivo systems. An in vitro, biosensor measurement takes place in a test tube, a culture dish, a microtiter plate or elsewhere outside a living organism. The sensor uses a bioreceptor and transducer as outlined above. An example of an in vitro biosensor is an enzyme-conductimetric biosensor for blood glucose monitoring. There is a challenge to create a biosensor that operates by the principle of point-of-care testing, i.e. at the location where the test is needed. Development of wearable biosensors is among such studies. The elimination of lab testing can save time and money. An application of a POCT biosensor can be for the testing of HIV in areas where it is difficult for patients to be tested. A biosensor can be sent directly to the location and a quick and easy test can be used.
An in vivo biosensor is an implantable device that operates inside the body. Of course, biosensor implants have to fulfill the strict regulations on sterilization in order to avoid an initial inflammatory response after implantation. The second concern relates to the long-term biocompatibility, i.e. the unharmful interaction with the body environment during the intended period of use. Another issue that arises is failure. If there is failure, the device must be removed and replaced, causing additional surgery. An example for application of an in vivo biosensor would be the insulin monitoring within the body, which is not available yet.
Most advanced biosensor implants have been developed for the continuous monitoring of glucose. The figure displays a device, for which a Ti casing and a battery as established for cardiovascular implants like pacemakers and defibrillators is used. Its size is determined by the battery as required for a lifetime of one year. Measured glucose data will be transmitted wirelessly out of the body within the MICS 402-405 MHz band as approved for medical implants.
Biosensors can also be integrated into mobile phone systems, making them user-friendly and accessible to a large number of users.
A common example of a commercial biosensor is the blood glucose biosensor, which uses the enzyme glucose oxidase to break blood glucose down. In doing so it first oxidizes glucose and uses two electrons to reduce the FAD (a component of the enzyme) to FADH2. This in turn is oxidized by the electrode in a number of steps. The resulting current is a measure of the concentration of glucose. In this case, the electrode is the transducer and the enzyme is the biologically active component.
A canary in a cage, as used by miners to warn of gas, could be considered a biosensor. Many of today's biosensor applications are similar, in that they use organisms which respond to toxic substances at a much lower concentrations than humans can detect to warn of their presence. Such devices can be used in environmental monitoring, trace gas detection and in water treatment facilities.
Since initial publication, IRIS has been adapted to perform various functions. First, IRIS integrated a fluorescence imaging capability into the interferometric imaging instrument as a potential way to address fluorescence protein microarray variability. Briefly, the variation in fluorescence microarrays mainly derives from inconsistent protein immobilization on surfaces and may cause misdiagnoses in allergy microarrays. To correct for any variation in protein immobilization, data acquired in the fluorescence modality is then normalized by the data acquired in the label-free modality. IRIS has also been adapted to perform single nanoparticle counting by simply switching the low magnification objective used for label-free biomass quantification to a higher objective magnification.C. A. Lopez, G. G. Daaboul, R. S. Vedula, E. Ozkumur, D. A. Bergstein, T. W. Geisbert, H. Fawcett, B. B. Goldberg, J. H. Connor, and M. S. Ünlü, "Label-free multiplexed virus detection using spectral reflectance imaging," Biosensors and Bioelectronics, 2011 This modality enables size discrimination in complex human biological samples. Monroe et al. used IRIS to quantify protein levels spiked into human whole blood and serum and determined allergen sensitization in characterized human blood samples using zero sample processing. Other practical uses of this device include virus and pathogen detection.
A range of immuno- and ligand-binding assays for the detection and measurement of small molecules such as water-soluble vitamins and chemical contaminants (drug residues) such as sulfonamides and Beta-agonists have been developed for use on SPR based sensor systems, often adapted from existing ELISA or other immunological assay. These are in widespread use across the food industry.
For example, bionanotechnologists developed a viable biosensor, , that can detect levels of diverse water pollution.
Deneb Karentz, a researcher at the Laboratory of Radio-biology and Environmental Health (University of California, San Francisco) has devised a simple method for measuring ultraviolet penetration and intensity. Working in the Antarctic Ocean, she submerged to various depths thin plastic bags containing special strains of E. coli that are almost totally unable to repair ultraviolet radiation damage to their DNA. Bacterial death rates in these bags were compared with rates in unexposed control bags of the same organism. The bacterial "biosensors" revealed constant significant ultraviolet damage at depths of 10 m and frequently at 20 and 30 m. Karentz plans additional studies of how ultraviolet may affect seasonal plankton Algal bloom (growth spurts) in the oceans.J. G. Black,"Principles and explorations", edition 5th.
Biological engineering researchers have created oncological biosensors for breast cancer. Breast cancer is the leading common cancer among women worldwide.Nordqvist, Christian. "Breast Cancer Cancer / Oncology Women's Health / Gynecology Breast Cancer: Causes, Symptoms and Treatments." Medical News Today. N.p., 5 May 2016. Web. An example would be a transferrin- quartz crystal microbalance (QCM). As a biosensor, quartz crystal microbalances produce oscillations in the frequency of the crystal's standing wave from an alternating potential to detect nano-gram mass changes. These biosensors are specifically designed to interact and have high selectivity for receptors on cell (cancerous and normal) surfaces. Ideally, this provides a quantitative detection of cells with this receptor per surface area instead of a qualitative picture detection given by mammograms.
Seda Atay, a biotechnology researcher at Hacettepe University, experimentally observed this specificity and selectivity between a QCM and MDA-MB 231 breast cells, MCF 7 cells, and starved MDA-MB 231 cells in vitro. With other researchers she devised a method of washing these different metastatic leveled cells over the sensors to measure mass shifts due to different quantities of transferrin receptors. Particularly, the metastatic power of breast cancer cells can be determined by Quartz crystal microbalances with nanoparticles and transferrin that would potentially attach to transferrin receptors on cancer cell surfaces. There is very high selectivity for transferrin receptors because they are over-expressed in cancer cells. If cells have high expression of transferrin receptors, which shows their high metastatic power, they have higher affinity and bind more to the QCM that measures the increase in mass. Depending on the magnitude of the nano-gram mass change, the metastatic power can be determined.
Additionally, in the last years, significant attentions have been focused to detect the biomarkers of lung cancer without biopsy. In this regard, biosensors are very attractive and applicable tools for providing rapid, sensitive, specific, stable, cost-effective and non-invasive detections for early lung cancer diagnosis. Thus, cancer biosensors consisting of specific biorecognition molecules such as antibodies, complementary nucleic acid probes or other immobilized biomolecules on a transducer surface. The biorecognition molecules interact specifically with the biomarkers (targets) and the generated biological responses are converted by the transducer into a measurable analytical signal. Depending on the type of biological response, various transducers are utilized in the fabrication of cancer biosensors such as electrochemical, optical and mass-based transducers.
Embedded biosensors for pathogenic signatures – such as of SARS-CoV-2 – that are wearable have been developed – such as face masks with built-in tests. See also: COVID-19 public transport R&D
New types of biosensor-chips could enable novel methods "such as drone-deployed pathogen sensors actively surveying air or wastewater". Protein-binding aptamers could be used for testing for infectious disease pathogens. Systems of (or robot skins) with built-in biosensors (or chemical sensors) and human-machine interfaces may enable wearable as well as remote sensing device- or robotic sensing of pathogens (as well as of several hazardous materials and tactile sensor).
Surface plasmon resonance sensors operate using a sensor chip consisting of a plastic cassette supporting a glass plate, one side of which is coated with a microscopic layer of gold. This side contacts the optical detection apparatus of the instrument. The opposite side is then contacted with a microfluidic flow system. The contact with the flow system creates channels across which reagents can be passed in solution. This side of the glass sensor chip can be modified in a number of ways, to allow easy attachment of molecules of interest. Normally it is coated in carboxymethyl dextran or similar compound.
The refractive index at the flow side of the chip surface has a direct influence on the behavior of the light reflected off the gold side. Binding to the flow side of the chip has an effect on the refractive index and in this way biological interactions can be measured to a high degree of sensitivity with some sort of energy. The refractive index of the medium near the surface changes when biomolecules attach to the surface, and the SPR angle varies as a function of this change.
Light of a fixed wavelength is reflected off the gold side of the chip at the angle of total internal reflection, and detected inside the instrument. The angle of incident light is varied in order to match the evanescent wave propagation rate with the propagation rate of the surface plasmon polaritons. This induces the evanescent wave to penetrate through the glass plate and some distance into the liquid flowing over the surface.
Other optical biosensors are mainly based on changes in absorbance or fluorescence of an appropriate indicator compound and do not need a total internal reflection geometry. For example, a fully operational prototype device detecting casein in milk has been fabricated. The device is based on detecting changes in absorption of a gold layer. A widely used research tool, the micro-array, can also be considered a biosensor.
Nanobiosensors use an immobilized bioreceptor probe that is selective for target analyte molecules. Nanomaterials are exquisitely sensitive chemical and biological sensors. Nanoscale materials demonstrate unique properties. Their large surface area to volume ratio can achieve rapid and low cost reactions, using a variety of designs.
Other evanescent wave biosensors have been commercialised using waveguides where the propagation constant through the waveguide is changed by the absorption of molecules to the waveguide surface. One such example, dual polarisation interferometry uses a buried waveguide as a reference against which the change in propagation constant is measured. Other configurations such as the Mach–Zehnder have reference arms lithographically defined on a substrate. Higher levels of integration can be achieved using resonator geometries where the resonant frequency of a ring resonator changes when molecules are absorbed.
Many techniques exist to detect DNA, which is usually a means to detect organisms that have that particular DNA. DNA sequences can also be used as described above. But more forward-looking approaches exist, where DNA can be synthesized to hold enzymes in a biological, stable gel. Other applications are the design of aptamers, sequences of DNA that have a specific shape to bind a desired molecule. The most innovative processes use DNA origami for this, creating sequences that fold in a predictable structure that is useful for detection.
Scientists have built prototype sensors to detect DNA of animals from sucked in air, "airborne eDNA".
"Nanoantennas" made out of DNA – a novel type of nano-scale optical antenna – can be attached to and produce a signal via fluorescence when these perform their biological functions, in particular for distinct conformational changes.
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