Biorecognition
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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.
Excessive use of pesticides could potentially harm the environment for a long time. The reason for this is that the banned pesticide is still likely to be used incorrectly. Carbofuran and other banned pesticides that remain in the environment may also have a negative effect on human beings. In order to provide a better chance for effective environmental screening, this thesis describes a prototype of a photometer tested with cholinesterase to potentially detect pesticides in the environment. The open-source portable photodetection platform uses a color-programmable red, green and blue light-emitting diode (RGB LED) as a light source and a TSL230R light frequency sensor. Acetylcholinesterase from Electrophorus electricus (AChE) with high similarity to human AChE was used for biorecognition. The Ellman method was selected as a standard method. Two analytical approaches were applied: (1) subtraction of the output values after a certain period of time and (2) comparison of the slope values of the linear trend. The optimal preincubation time for carbofuran with AChE was 7 min. The limits of detection for carbofuran were 6.3 nmol/L for the kinetic assay and 13.5 nmol/L for the endpoint assay. The paper demonstrates that the open alternative for commercial photometry is equivalent. The concept based on the OS3P/OS3P could be used as a large-scale screening system.
- MeSH
- acetylcholinesterasa MeSH
- cholinesterasové inhibitory MeSH
- cholinesterasy MeSH
- karbofuran * MeSH
- lidé MeSH
- pesticidy * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
This paper summarizes the area of biomedicinal polymers, which serve as nanomedicines even though they do not contain any anticancer or antiinflammatory drugs. These polymer nanomedicines with unique design are in the literature highlighted as a novel class of therapeutics called "drug-free macromolecular therapeutics." Their therapeutic efficacy is based on the tailored multiple presentations of biologically active vectors, i.e., peptides, oligopeptides, or oligosaccharides. Thus, they enable, for example, to directly induce the apoptosis of malignant cells by the crosslinking of surface slowly internalizing receptors, or to deplete the efficacy of tumor-associated proteins. The precise biorecognition of natural binding motifs by multiple vectors on the polymer construct remains the crucial part in the designing of these drug-free nanomedicines. Here, the rationales, designs, synthetic approaches, and therapeutic potential of drug-free macromolecular therapeutics consisting of various active vectors are described in detail. Recent developments and achievements for namely B-cell lymphoma treatment, Gal-3-positive tumors, inflammative liver injury, and bacterial treatment are reviewed and highlighted. Finally, a possible future prospect within this highly exciting new field of nanomedicine research is presented.
- MeSH
- antibakteriální látky farmakologie terapeutické užití MeSH
- antiflogistika farmakologie terapeutické užití MeSH
- lékové transportní systémy metody MeSH
- lidé MeSH
- makromolekulární látky chemie MeSH
- nádory * farmakoterapie MeSH
- nanomedicína metody MeSH
- polymery chemie MeSH
- protinádorové látky * farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy 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.
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.
An amperometric biosensor compatible with a flow injection analysis (FIA) for highly selective determination of acetaminophen (APAP) in a sample of human urine was developed. This biosensor is also suitable for use in the routine pharmaceutical practice. To prove this statement, two different commercially available pharmaceutical formulations were analyzed. This nano-(bio)electroanalytical device was made from a commercially available screen-printed carbon electrode covered by a thin layer of non-functionalized graphene (NFG) as amperometric transducer. A biorecognition layer was prepared from mushroom (Agaricus bisporus) tyrosinase (EC 1.14.18.1) cross-linked using glutaraldehyde, where resulting aggregates were covered by Nafion®, a known ion exchange membrane. Owing to the use of tyrosinase and presence of NFG, the developed analytical instrument is able to measure even at potentials of 0 V. Linear ranges differ according to choice of detection potential, namely up to 130 μmol L-1 at 0 V, up to 90 μmol L-1 at -0.1 V, and up to 70 μmol L-1 at -0.15 V. The first mentioned linear range is described by the equation Ip [μA] = 0.236 - 0.1984c [μmol L-1] and correlation coefficient r = 0.9987; this equation was used to quantify the content of APAP in each sample. The limit of detection of APAP was estimated to be 1.1 μmol L-1. A recovery of 96.8% (c = 25 μmol L-1, n = 5 measurements) was calculated. The obtained results show that FIA is a very selective method for APAP determination, being comparable to the chosen reference method of reversed-phase high-performance liquid chromatography.
- MeSH
- Agaricus enzymologie MeSH
- analýza moči přístrojové vybavení metody MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- design vybavení MeSH
- lidé MeSH
- limita detekce MeSH
- neopioidní analgetika moč MeSH
- paracetamol moč MeSH
- průtoková injekční analýza přístrojové vybavení metody MeSH
- tyrosinasa chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
Analytical devices that combine sensitive biological component with a physicochemical detector hold a great potential for various applications, e.g., environmental monitoring, food analysis or medical diagnostics. Continuous efforts to develop inexpensive sensitive biodevices for detecting target substances typically focus on the design of biorecognition elements and their physical implementation, while the methods for processing signals generated by such devices have received far less attention. Here, we present fundamental considerations related to signal processing in biosensor design and investigate how undemanding signal treatment facilitates calibration and operation of enzyme-based biodevices. Our signal treatment approach was thoroughly validated with two model systems: (i) a biodevice for detecting chemical warfare agents and environmental pollutants based on the activity of haloalkane dehalogenase, with the sensitive range for bis(2-chloroethyl) ether of 0.01-0.8 mM and (ii) a biodevice for detecting hazardous pesticides based on the activity of γ-hexachlorocyclohexane dehydrochlorinase with the sensitive range for γ-hexachlorocyclohexane of 0.01-0.3 mM. We demonstrate that the advanced signal processing based on curve fitting enables precise quantification of parameters important for sensitive operation of enzyme-based biodevices, including: (i) automated exclusion of signal regions with substantial noise, (ii) derivation of calibration curves with significantly reduced error, (iii) shortening of the detection time, and (iv) reliable extrapolation of the signal to the initial conditions. The presented simple signal curve fitting supports rational design of optimal system setup by explicit and flexible quantification of its properties and will find a broad use in the development of sensitive and robust biodevices.
- MeSH
- biosenzitivní techniky metody MeSH
- chemické bojové látky analýza MeSH
- chlorované uhlovodíky analýza MeSH
- enzymy metabolismus MeSH
- ether analogy a deriváty analýza MeSH
- hexany analýza MeSH
- hydrolasy metabolismus MeSH
- kalibrace MeSH
- látky znečišťující životní prostředí analýza MeSH
- lyasy metabolismus MeSH
- počítačové zpracování signálu * MeSH
- senzitivita a specificita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Early diagnosis of ongoing malignant disease is crucial to improve survival rate and life quality of the patients and requires sensitive detection of specific biomarkers e.g. prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), etc. In spite of current technological advances, malignant diseases are still identified in rather late stages, which have detrimental effect on the prognosis and treatment of the disease. Here, we present a biosensor able to detect fetuin-A, a potential multibiomarker. The biosensing platform is based on polymer brush combining antifouling monomer units of N-(2-hydroxypropyl)methacrylamide (HPMA) and carboxybetaine methacrylamide (CBMAA), statistically copolymerized by surfaceinitiated atom transfer radical polymerization. The copolymer poly(HPMA-co-CBMAA) exhibits excellent non-fouling properties in the most relevant biological media (i.e. blood plasma) as well as antithrombogenic surface properties by preventing the adhesion of blood components (i.e. leukocytes; platelets; and erythrocytes). Moreover, the polymer brush can be easily functionalized with biorecognition elements maintaining high resistance to blood fouling and the binding capacity can be regulated by tuning the ratio between CBMAA and HPMA units. The superior antifouling properties of the copolymer even after biofunctionalization were exploited to fabricate a new plasmonic biosensor for the analysis of fetuin-A in real clinical blood plasma samples. The assay used in this work can be explored as labelfree affinity biosensor for diagnostics of different biomarkers in real clinical plasma samples and to shift the early biomarker detection toward novel biosensor technologies allowing point of care analysis.
- MeSH
- biologické markery krev MeSH
- biosenzitivní techniky metody MeSH
- fetuin A analýza metabolismus MeSH
- lidé MeSH
- povrchová plasmonová rezonance metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
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
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
