Biorecognition element
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Antibodies are macromolecular structures with a high affinity to a molecule called the antigen. Though the natural aim of antibodies is the protection of the body from pathogens, they have been found to play a useful part in a number of specific treatments and diagnoses. This wider role for antibodies extends to assays where antibodies provide a recognition capability improving available physical methods. Photometrical methods such as ELISA or flow cytometry and electrochemical, optical or piezoelectric immunosensors (biosensors) are typical examples. The present review summarizes facts about the parameters and the production of antibodies. The structure of immunoglobulins, and the production and purification of monoclonal and polyclonal antibodies are described in four chapters. The review was written in order to collect the available knowledge on antibodies into one study improving orientation in this field for anyone wanting to construct immunoassays.
- MeSH
- adjuvancia imunologická terapeutické užití MeSH
- biologické bojové látky MeSH
- biosenzitivní techniky metody trendy využití MeSH
- chemické bojové látky izolace a purifikace MeSH
- ELISA metody trendy využití MeSH
- financování organizované MeSH
- imunoanalýza metody trendy využití MeSH
- imunoglobuliny diagnostické užití imunologie izolace a purifikace MeSH
- laboratorní zvířata MeSH
- lidé MeSH
- monoklonální protilátky diagnostické užití imunologie izolace a purifikace MeSH
- protilátky diagnostické užití imunologie izolace a purifikace MeSH
- tvorba protilátek MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
The cholinesterase biosensors are analytical devices suitable for assay of cholinesterase inhibitors in the course of suppression of the biorecognition element activity. Both acetylcholinesterase and butyrylcholinesterase are used for their assay. The biosensors can be used for assay of organophosphorus and carbamate pesticides, nerve agents, drugs in Alzheimer´s disease and myasthenia treatment. The review summarizes the knowledge of cholinesterases, their immobilization and inhibitors as well as shows examples of their activities.
- MeSH
- acetylcholinesterasa MeSH
- Alzheimerova nemoc MeSH
- biosenzitivní techniky * metody využití MeSH
- butyrylcholinesterasa MeSH
- cholinesterasové inhibitory * MeSH
- dichlorvos * analýza normy MeSH
- karbamáty * farmakologie MeSH
- lidé MeSH
- organofluorofosfonáty * farmakologie MeSH
- reaktivátory cholinesterasy analýza MeSH
- sarin farmakologie MeSH
- Check Tag
- lidé MeSH
Recent advances in optical sensing technologies underpin the development of high-performance, surface-sensitive analytical tools capable of reliable and precise detection of molecular targets in complex biological media in non-laboratory settings. Optical fibre sensors guide light to and from a region of interest, enabling sensitive measurements of localized environments. This positions optical fibre sensors as a highly promising technology for a wide range of biochemical and healthcare applications. However, their performance in real-world biological media is often limited by the absence of robust post-modification strategies that provide both high biorecognition and antifouling capabilities. In this study, we present the proof-of-concept antifouling and biorecognition performance of a polymer brush nano-coating synthesized at the sensing region of optical fibre long-period grating (LPG) sensors. Using a newly developed antifouling terpolymer brush (ATB) composed of carboxybetaine methacrylamide, sulfobetaine methacrylamide, and N-(2-hydroxypropyl)methacrylamide, we achieve state-of-the-art antifouling properties. The successful on-fibre ATB synthesis is confirmed through scanning electron microscopy (SEM), fluorescence microscopy, and label-free bio-detection experiments based on antibody-functionalized ATB-coated LPG optical fibres. Despite the challenges in handling optical fibres during polymerization, the resulting nano-coating retains its remarkable antifouling properties upon exposure to blood plasma and enables biorecognition element functionalization. These capabilities are demonstrated through the detection of IgG in buffer and diluted blood plasma using anti-IgG-functionalized ATB-coated sensing regions of LPG fibres in both label-based (fluorescence) and label-free real-time detection experiments. The results show the potential of ATB-coated LPG fibres for use in analytical biosensing applications.
A surface plasmon resonance (SPR) biosensor for simultaneous detection of multiple organic pollutants exhibiting endocrine-disrupting activity, namely atrazine, benzo[a]pyrene, 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-nonylphenol, is reported. The biosensor utilizes a multichannel SPR sensor based on wavelength modulation of SPR and wavelength division multiplexing (WDM) of sensing channels, antibodies as biorecognition element and a competitive immunoassay detection format. An analysis time of 45 min (including 30-min incubation of the sample with antibodies) and limits of detection as low as 0.05, 0.07, 0.16 and 0.26 ng mL(-1) are demonstrated for benzo[a]pyrene, atrazine, 2,4-D and 4-nonylphenol, respectively. The biosensor is also shown to be regenerable and suitable for repeated use.
- MeSH
- atrazin analýza MeSH
- benzopyren analýza MeSH
- biosenzitivní techniky metody přístrojové vybavení MeSH
- časové faktory MeSH
- endokrinní disruptory analýza MeSH
- fenoly analýza MeSH
- financování organizované MeSH
- imunoanalýza metody MeSH
- kyselina 2,4-dichlorfenoxyoctová analýza MeSH
- povrchová plasmonová rezonance metody přístrojové vybavení MeSH
- senzitivita a specificita MeSH
... Biorecognition elements and their immobilization 9 -- 3.3. ...
32 s. : il. ; 21 cm
- MeSH
- biofyzika MeSH
- biosenzitivní techniky MeSH
- mikrochemie MeSH
- optické jevy MeSH
- povrchová plasmonová rezonance MeSH
- Publikační typ
- vysokoškolské kvalifikační práce MeSH
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- fyzika, biofyzika
Medical diagnostics aims at specific localization of molecular targets as well as detection of abnormalities associated with numerous diseases. Molecularly imprinted polymers (MIPs) represent an approach of creating a synthetic material exhibiting selective recognition properties toward the desired template. The fabricated target-specific MIPs are usually well reproducible, economically efficient, and stable under critical conditions as compared to routinely used biorecognition elements such as fluorescent proteins, antibodies, enzymes, or aptamers and can even be created to those targets for which no antibodies are available. In this review, we summarize the methods of polymer fabrication. Further, we provide key for selection of the core material with imaging function depending on the imaging modality used. Finally, MIP-based imaging applications are highlighted and presented in a comprehensive form from different aspects. STATEMENT OF SIGNIFICANCE: In this review, we summarize the methods of polymer fabrication. Key applications of Molecularly imprinted polymers (MIPs) in imaging are highlighted and discussed with regard to the selection of the core material for imaging as well as commonly used imaging targets. MIPs represent an approach of creating a synthetic material exhibiting selective recognition properties toward the desired template. The fabricated target-specific MIPs are usually well reproducible, economically efficient, and stable under critical conditions as compared to routinely used biorecognition elements, e.g., antibodies, fluorescent proteins, enzymes, or aptamers, and can even be created to those targets for which no antibodies are available.
Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in many heart pathologies. Biosensing of these parameters represents an important tool in drug development and disease modeling. Human embryonic stem cells and especially patient specific induced pluripotent stem cell-derived cardiomyocytes are well established as cardiac disease model.. Here, a live stem cell derived embryoid body (EB) based cardiac cell syncytium served as a biorecognition element coupled to the microcantilever probe from atomic force microscope thus providing reliable micromechanical cellular biosensor suitable for whole-day testing. The biosensor was optimized regarding the type of cantilever, temperature and exchange of media; in combination with standardized protocol, it allowed testing of compounds and conditions affecting the biomechanical properties of EB. The studied effectors included calcium , drugs modulating the catecholaminergic fight-or-flight stress response such as the beta-adrenergic blocker metoprolol and the beta-adrenergic agonist isoproterenol. Arrhythmogenic effects were studied using caffeine. Furthermore, with EBs originating from patient's stem cells, this biosensor can help to characterize heart diseases such as dystrophies.
- MeSH
- agonisté adrenergních beta-receptorů farmakologie MeSH
- antagonisté beta-1-adrenergních receptorů farmakologie MeSH
- biomechanika účinky léků MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- buněčné kultury přístrojové vybavení metody MeSH
- buněčné linie MeSH
- design vybavení MeSH
- isoprenalin farmakologie MeSH
- kardiomyocyty cytologie účinky léků metabolismus MeSH
- kontrakce myokardu účinky léků MeSH
- lidé MeSH
- metoprolol farmakologie MeSH
- mikroskopie atomárních sil přístrojové vybavení metody MeSH
- pluripotentní kmenové buňky cytologie MeSH
- preklinické hodnocení léčiv přístrojové vybavení metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Proteins are generally detected as biomarkers for tracing or determining various disorders in organisms. Biomarker proteins can be tracked in samples with various origins and in different concentrations, revealing whether an organism is in a healthy or unhealthy state. In regard to detection, electrochemical biosensors are a potential fusion of electronics, chemistry, and biology, allowing for fast and early point-of-care detection from a biological sample with the advantages of high sensitivity, simple construction, and easy operation. Peptides present a promising approach as a biorecognition element when connected with electrochemical biosensors. The benefits of short peptides lie mainly in their good stability and selective affinity to a target analyte. Therefore, peptide-based electrochemical biosensors (PBEBs) represent an alternative approach for the detection of different protein biomarkers. This review provides a summary of the past decade of recently proposed PBEBs designed for protein detection, dividing them according to different protein types: (i) enzyme detection, including proteases and kinases; (ii) antibody detection; and (iii) other protein detection. According to these protein types, different sensing mechanisms are discussed, such as the peptide cleavage by a proteases, phosphorylation by kinases, presence of antibodies, and exploiting of affinities; furthermore, measurements are obtained by different electrochemical methods. A discussion and comparison of various constructions, modifications, immobilization strategies and different sensing techniques in terms of high sensitivity, selectivity, repeatability, and potential for practical application are presented.
DNA methylation plays an important role in physiological and pathological processes. Several genetic diseases and most malignancies tend to be associated with aberrant DNA methylation. Among other analytical methods, electrochemical approaches have been successfully employed for characterisation of DNA methylation patterns that are essential for the diagnosis and treatment of particular diseases. This article discusses current trends in the electrochemical sensing and biosensing of DNA methylation. Particularly, it provides an overview of applied electrode materials, electrode modifications and biorecognition elements applications with an emphasis on strategies that form the core DNA methylation detection approaches. The three main strategies as (i) bisulfite treatment, (ii) cleavage by restriction endonucleases, and (iii) immuno/affinity reaction were described in greater detail. Additionally, the availability of the reviewed platforms for early cancer diagnosis and the approval of methylation inhibitors for anticancer therapy were discussed.
- MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- DNA analýza genetika MeSH
- elektrochemické techniky přístrojové vybavení metody MeSH
- elektrody MeSH
- lidé MeSH
- metylace DNA * MeSH
- nádory diagnóza genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Biosensors consist of a biological entity that recognizes the target analyte and the transducer that translates the biorecognition event into an electrical signal. The use of biological materials as recognizing elements imparts to biosensors the ability to specifically respond to the analyte of interest, distinguishing it from structurally similar compounds. Amperometric biosensors are based on measurements of the electric current resulting from the oxidation or reduction of an electroactive species, by keeping a constant potential at the working electrode. Enzymes can be immobilized in a thin layer on the transducer surface. Besides the reactive layer or membrane many biosensors contain one or several inner or outer membranes with different functions. The protective inner membrane may decrease the influence of interfering species. The outer membrane serves as a diffusion-limiting barrier to the enzyme substrate. Biosensors are considered biocompatible if their implantation does not affect normal functioning of the host medium and, vice versa, the host medium does not materially affect their normal operation. The present review focuses on the immobilisation techniques and preparation and utilization of protective inner and outer diffusionlimiting membranes of amperometric biosensors.
