RESUMO
Biosensors are advanced devices for analysis of composition of blood, urine, environmental samples, and many other media. Their current development is tightly linked with nanomaterials, such as zeolites and zeolitic imidazolate framework (ZIFs). The present review describes electrochemical (amperometric, conductometric, ISFET) and optical (fluorescent and colorimetric) biosensors that incorporate zeolites and ZIFs in their biorecognition elements. The biosensors are based on immobilized enzymes (such as glucose oxidase, urease, and acetylcholinesterase), antibodies, DNA, and aptamers. The review present reasons for application of these nanomaterials, and discusses advantages of zeolite- and ZIF-containing biosensors over other biosensors. In most cases, the biosensors have improved sensitivity, better limit of detection, wider linear range, and other improved characteristics. It is demonstrated that immobilization of biomolecules such as enzymes or antibodies on the surface of zeolites and ZIFs enables creation of unique advanced biosensors that have a potential for further development and practical applications.
Assuntos
Técnicas Biossensoriais , Zeolitas , Zeolitas/química , Acetilcolinesterase , Enzimas Imobilizadas/química , Glucose OxidaseRESUMO
In the review, the principles and main purposes of using multienzyme systems in electrochemical biosensors are analyzed. Coupling several enzymes allows an extension of the spectrum of detectable substances, an increase in the biosensor sensitivity (in some cases, by several orders of magnitude), and an improvement of the biosensor selectivity, as showed on the examples of amperometric, potentiometric, and conductometric biosensors. The biosensors based on cascade, cyclic and competitive enzyme systems are described alongside principles of function, advantages, disadvantages and practical use for real sample analyses in various application areas (food production and quality control, clinical diagnostics, environmental monitoring). The complications and restrictions regarding the development of multienzyme biosensors are evaluated. The recommendations on the reasonability of elaboration of novel multienzyme biosensors are given.
Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Peroxidase do Rábano Silvestre/metabolismo , NADPH Desidrogenase/metabolismo , Carboidratos/análise , Peroxidase do Rábano Silvestre/química , Humanos , Lipídeos/análise , NADPH Desidrogenase/químicaRESUMO
The work was aimed at the development of a biosensor array for the simultaneous determination of six solutes (glutamate, glucose, choline, acetylcholine, lactate, and pyruvate) in aqueous solutions. Enzymes selective for these substrates were immobilized on the surface of amperometric platinum disc electrodes and served as bioselective elements of a biosensor array. Direct enzymatic analysis by the developed biosensors provided high sensitivity to the tested substrates (limits of detection were 1-5⯵M). The linear ranges of the biosensors were from 0.001-0.01â¯mM to 0.2-2.5â¯mM. The influence of solution pH, ionic strength and buffer capacity on the biosensor responses was investigated; the conditions for simultaneous operation of all the bioselective elements were optimized. The absence of any cross-influence of the substrates of enzymatic systems used was shown as well as a high selectivity of the biosensors and the absence of any impact of interfering substances (ascorbic acid, dopamine, cysteine, paracetamol). The developed biosensor array had good response reproducibility and storage stability. The array is suitable for rapid (0.5-1â¯min) and simple simultaneous determination of glutamate, glucose, choline, acetylcholine, lactate, and pyruvate in aqueous (biological) samples; furthermore, the creation of a single chip with six sensitive elements is possible as well as the addition of other biosensors.
Assuntos
Acetilcolina/análise , Técnicas Biossensoriais , Colina/análise , Técnicas Eletroquímicas/instrumentação , Eletrodos , Enzimas Imobilizadas/química , Glucose/análise , Ácido Glutâmico/análise , Ácido Láctico/análise , Ácido Pirúvico/análise , Soluções Tampão , Concentração de Íons de Hidrogênio , Limite de Detecção , Concentração Osmolar , Reprodutibilidade dos TestesRESUMO
Electrochemical enzyme-based biosensors are one of the largest and commercially successful groups of biosensors. Integration of nanomaterials in the biosensors results in significant improvement of biosensor sensitivity, limit of detection, stability, response rate and other analytical characteristics. Thus, new functional nanomaterials are key components of numerous biosensors. However, due to the great variety of available nanomaterials, they should be carefully selected according to the desired effects. The present review covers the recent applications of various types of nanomaterials in electrochemical enzyme-based biosensors for the detection of small biomolecules, environmental pollutants, food contaminants, and clinical biomarkers. Benefits and limitations of using nanomaterials for analytical purposes are discussed. Furthermore, we highlight specific properties of different nanomaterials, which are relevant to electrochemical biosensors. The review is structured according to the types of nanomaterials. We describe the application of inorganic nanomaterials, such as gold nanoparticles (AuNPs), platinum nanoparticles (PtNPs), silver nanoparticles (AgNPs), and palladium nanoparticles (PdNPs), zeolites, inorganic quantum dots, and organic nanomaterials, such as single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs), carbon and graphene quantum dots, graphene, fullerenes, and calixarenes. Usage of composite nanomaterials is also presented.
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A new conductometric biosensor based on coimmobilized urease and arginase has been developed for arginine determination in pharmaceutics. First, the main parameters of the selected method of immobilization (concentrations of arginase, urease, and glutaraldehyde, time of incubation) were optimized. An influence of the solution parameters (buffer ionic strength, capacity, pH, Mn2+ concentration) on the biosensor operation was studied, working conditions were optimized. After biosensor optimization, the main analytical characteristics were as follows. The limit of detection - 2.5⯵M, the linear range - 2.5-500⯵M, the sensitivity to arginine 13.4⯱â¯2.4⯵S/mM, the response time - 20â¯s. The signals repeatability and operational stability in continuous exploitation were studied over one working day and during one week. Additionally, the selectivity of the developed biosensor towards arginine was essayed relative to other amino acids. The developed biosensor has been used to measure arginine concentrations in some drugs. The results obtained were in high correlation with the characteristics declared by producers.
Assuntos
Arginina/análise , Técnicas Biossensoriais , Condutometria/instrumentação , Preparações Farmacêuticas/química , Arginase/metabolismo , Enzimas Imobilizadas/metabolismo , Limite de Detecção , Membranas Artificiais , Concentração Osmolar , Reprodutibilidade dos Testes , Urease/metabolismoRESUMO
In this work, we studied the conditions of deposition of a semipermeable polyphenylenediamine (PPD)-based membrane on amperometric disk platinum electrodes. Restricting an access of interfering substances to the electrode surface, the membrane prevents their impact on the sensor operation. Two methods of membrane deposition by electropolymerization were compared-at varying potential (cyclic voltammetry) and at constant potential. The cyclic voltammetry was shown to be easier in performing and providing better properties of the membrane. The dependence of PPD membrane effectiveness on the number of cyclic voltammograms and phenylenediamine concentration was analyzed. It was shown that the impact of interfering substances (ascorbic acid, dopamine, cysteine, uric acid) on sensor operation could be completely avoided using three cyclic voltammograms in 30 mM phenylenediamine. On the other hand, when working with diluted samples, i.e., at lower concentrations of electroactive substances, it is reasonable to decrease the phenylenediamine concentration to 5 mM, which would result in a higher sensitivity of transducers to hydrogen peroxide due to a thinner PPD layer. The PPD membrane was tested during continuous operation and at 8-day storage and turned out to be efficient in sensor and biosensors.
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In this work, we developed a new amperometric biosensor for glutamate detection using a typical method of glutamate oxidase (GlOx) immobilization via adsorption on silicalite particles. The disc platinum electrode (d = 0.4 mm) was used as the amperometric sensor. The procedure of biosensor preparation was optimized. The main parameters of modifying amperometric transducers with a silicalite layer were determined along with the procedure of GlOx adsorption on this layer. The biosensors based on GlOx adsorbed on silicalite demonstrated high sensitivity to glutamate. The linear range of detection was from 2.5 to 450 µM, and the limit of glutamate detection was 1 µM. It was shown that the proposed biosensors were characterized by good response reproducibility during hours of continuous work and operational stability for several days. The developed biosensors could be applied for determination of glutamate in real samples.
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The paper presents a simple and inexpensive reusable biosensor for determination of the concentration of adenosine-5'-triphosphate (ATP) in aqueous samples. The biosensor is based on a conductometric transducer which contains two pairs of gold interdigitated electrodes. An enzyme hexokinase was immobilized onto one pair of electrodes, and bovine serum albumin-onto another pair (thus, a differential mode of measurement was used). Conditions of hexokinase immobilization on the transducer by cross-linking via glutaraldehyde were optimized. Influence of experimental conditions (concentration of magnesium ions, ionic strength and concentration of the working buffer) on the biosensor work was studied. The reproducibility of biosensor responses and operational stability of the biosensor were checked during one week. Dry storage at -18 °C was shown to be the best conditions to store the biosensor. The biosensor was successfully applied for measurements of ATP concentration in pharmaceutical samples. The proposed biosensor may be used in future for determination of ATP and/or glucose in water samples.
Assuntos
Trifosfato de Adenosina/análise , Técnicas Biossensoriais/métodos , Condutometria/métodos , Hexoquinase/química , Trifosfato de Adenosina/química , Animais , Técnicas Biossensoriais/instrumentação , Soluções Tampão , Condutometria/instrumentação , Eletrodos , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Glutaral/química , Ouro/química , Hexoquinase/metabolismo , Magnésio/química , Concentração Osmolar , Saccharomyces cerevisiae/enzimologia , TransdutoresRESUMO
The conductometric sensor based on 25,27-di-(5-thio-octyloxy)calix[4]arene-crown-6 was developed for the quantitative analysis of ammonium. The calixarene was immobilized on the surface of the planar interdigitated electrodes by attachment of its dialkyl sulfide groups to the surface of the gold electrodes. The intrinsic ability of the calixarene to capture ammonium was studied in the conductometric measuring mode and by the electrochemical impedance spectroscopy. The developed sensor showed high selectivity to ammonium in the presence of mono-, di-, and trivalent cations. Selective and highly sensitive detection of ammonium resulted from the complexation between the ammonium ions and a crown-ether fragment of the upper rim of the 25,27-di-(5-thio-octyloxy)calix[4]arene-crown-6 macrocycle. The developed sensor had high signal repeatability. Its sensitivity was found to be satisfactory for the forthcoming sensor application in the water-sample analysis; the linear range was 0.01-1.5 mM and limit of detection 10 µM.
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Development of a conductometric biosensor for the urea detection has been reported. It was created using a non-typical method of the recombinant urease immobilization via adsorption on nanoporous particles of silicalite. It should be noted that this biosensor has a number of advantages, such as simple and fast performance, the absence of toxic compounds during biosensor preparation, and high reproducibility (RSD = 5.1 %). The linear range of urea determination by using the biosensor was 0.05-15 mM, and a lower limit of urea detection was 20 µM. The bioselective element was found to be stable for 19 days. The characteristics of recombinant urease-based biomembranes, such as dependence of responses on the protein and ion concentrations, were investigated. It is shown that the developed biosensor can be successfully used for the urea analysis during renal dialysis.
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This report describes technical improvements to the manufacture of a carbon fibre electrode for the stable and sensitive detection of H2O2 (detection limit at 0.5 microM). This electrode was also modified through the co-immobilisation of acetylcholinesterase (AChE) and/or choline oxidase (ChOx) in a bovine serum albumin (BSA) membrane for the development of a sensor for in vivo measurements of acetylcholine and choline. Amperometric measurements were performed using a conventional three-electrode system forming part of a flow-injection set-up at an applied potential of 800-1100 mV relative to an Ag/AgCl reference electrode. The optimised biosensor obtained was reproducible and stable, and exhibited a detection limit of 1 microM for both acetylcholine and choline. However, due to the high operating potential used, the biosensor was prone to substantial interference from other electroactive compounds, such as ascorbic acid. Therefore, in a further step, a mediated electron transfer approach was used that incorporated horseradish peroxidase into an osmium-based redox hydrogel layered onto the active surface of the electrode. Afterwards, a Nafion layer and a coating containing AChE and/or ChOx co-immobilised in a BSA membrane were successively deposited. This procedure further increased the selectivity of the biosensor, when operated in the same flow-injection system but at an applied potential of -50 mV relative to an Ag/AgCl reference electrode. The sensor exhibited good selectivity and a high sensitivity over a concentration range (0.3-100 microM) suitable for the measurement of choline and acetylcholine in vivo.
Assuntos
Acetilcolina/análise , Técnicas Biossensoriais/instrumentação , Carbono , Colina/análise , Acetilcolinesterase , Oxirredutases do Álcool , Química Encefálica , Fibra de Carbono , Peroxidase do Rábano Silvestre , Humanos , MicroeletrodosRESUMO
This study was aimed at the development of a conductometric biosensor based on acetylcholinesterase considering the feasibility of its application for the inhibitory analysis of various toxicants. In this paper, the optimum conditions for enzyme immobilization on the transducer surface are selected as well as the optimum concentration of substrate for inhibitory analysis. Sensitivity of the developed biosensor to different classes of toxic compounds (organophosphorus pesticides, heavy metal ions, surfactants, aflatoxin, glycoalkaloids) was tested. It is shown that the developed biosensor can be successfully used for the analysis of pesticides and mycotoxins, as well as for determination of total toxicity of the samples. A new method of biosensor analysis of toxic substances of different classes in complex multicomponent aqueous samples is proposed.
Assuntos
Técnicas Biossensoriais , Eletroquímica/métodos , Substâncias Perigosas/toxicidade , Estudos de ViabilidadeRESUMO
Creatine kinase (CK: adenosine-5-triphosphate-creatine phosphotransferase) is an important enzyme of muscle cells; the presence of a large amount of the enzyme in blood serum is a biomarker of muscular injuries, such as acute myocardial infarction. This work describes a bi-enzyme (glucose oxidase and hexokinase based) biosensor for rapid and convenient determination of CK activity by measuring the rate of ATP production by this enzyme. Simultaneously the biosensor determines glucose concentration in the sample. Platinum disk electrodes were used as amperometric transducers. Glucose oxidase and hexokinase were co-immobilized via cross-linking with BSA by glutaraldehyde and served as a biorecognition element of the biosensor. The biosensor work at different concentrations of CK substrates (ADP and creatine phosphate) was investigated; optimal concentration of ADP was 1mM, and creatine phosphate - 10 mM. The reproducibility of the biosensor responses to glucose, ATP and CK during a day was tested (relative standard deviation of 15 responses to glucose was 2%, to ATP - 6%, to CK - 7-18% depending on concentration of the CK). Total time of CK analysis was 10 min. The measurements of creatine kinase in blood serum samples were carried out (at 20-fold sample dilution). Twentyfold dilution of serum samples was chosen as optimal for CK determination. The biosensor could distinguish healthy and ill people and evaluate the level of CK increase. Thus, the biosensor can be used as a test-system for CK analysis in blood serum or serve as a component of multibiosensors for determination of important blood substances. Determination of activity of other kinases by the developed biosensor is also possible for research purposes.
Assuntos
Técnicas Biossensoriais , Creatina Quinase/sangue , Enzimas Imobilizadas/metabolismo , Glucose Oxidase/metabolismo , Hexoquinase/metabolismo , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Técnicas Eletroquímicas , Glutaral/metabolismo , Fosfocreatina/metabolismo , Soroalbumina Bovina/metabolismoRESUMO
This study aimed at the development and optimization of a potentiometric biosensor based on pH-sensitive field-effect transistors and acetylcholinesterase for aflatoxin B1 determination in real samples. Optimal conditions for bioselective elements operation were defined and analytical characteristics of the proposed biosensor were studied. The proposed biosensor characterized high operational stability and reproducibility of signal. Selectivity of acetylcholinesterase-biosensor to aflatoxins in relation to other groups of toxic substances was analyzed. The developed biosensor was applied to the determination of aflatoxin B1 in real samples (sesame, walnut and pea).
Assuntos
Acetilcolinesterase/metabolismo , Aflatoxina B1/análise , Técnicas Biossensoriais/instrumentação , Transistores Eletrônicos , Animais , Concentração de Íons de Hidrogênio , PotenciometriaRESUMO
This article describes a biosensor based on pH-sensitive field-effect transistors (pH-FETs) as transducer, and immobilised enzyme tyrosinase as biorecognition element, which was used for the determination of phenolic compounds in water solutions. The biologically active membrane was formed by cross-linking of tyrosinase with bovine serum albumin (BSA) in saturated glutaraldehyde (GA) vapours on the sensitive transducer surface. The main analytical characteristics were studied under different conditions as well as the possibility to optimise these working parameters. Different factors such as the pH of immobilisation, the enzyme loading, the time of exposition to glutaraldehyde vapours were investigated in regards to the influence on sensitivity, limit of detection, dynamic range, and operational and storage stability.
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Técnicas Biossensoriais , Concentração de Íons de Hidrogênio , Fenóis/análise , Enzimas Imobilizadas/química , Monofenol Mono-Oxigenase/química , Potenciometria , Sensibilidade e EspecificidadeRESUMO
In this work a novel biosensor for arginine determination based on the urease inhibition effect has been proposed. Ion-selective field effect transistors were used as transducers. Urease immobilized in glutaraldehyde vapor served as a biorecognition element of the biosensor. Significant part of the work was aimed at proving the urease inhibition by arginine. Optimal concentration of urea for arginine determination was chosen. Detection limit for arginine was 0.05 mM. The biosensor selectivity towards different amino acids was studied. The results of quantitative determination of l-arginine in the real sample (a drinkable solution "Arginine Veyron") were in good agreement with the producer's data (a relative error was 5.2%). The biosensor showed a good reproducibility of arginine determination.
Assuntos
Arginina/análise , Técnicas Biossensoriais , Urease/química , Animais , Bovinos , Limite de DetecçãoRESUMO
Urea biosensor based on zeolite-adsorbed urease was applied for analysis of blood serum samples. It should be noted, that this biosensor has a number of advantages, such as simple and fast performance, the absence of toxic compounds during biosensor preparation, high reproducibility and repeatability (RSD=9% and 4%, respectively). The linear range of urea determination by using the biosensor was 0.003-0.75 mM, and the limit of urea detection was 3 µM. The method of standard addition was used for analysis of serum samples with 500-fold dilution. Total time of analysis was 10 min. Good reproducibility of urea determination in real samples was demonstrated (RSD=10%). Biosensor results were verified by using a common method of urea determination (diacetyl monoxime reaction). It was shown that by using this biosensor distinguishing healthy people from people with renal dysfunction becomes easier.
Assuntos
Técnicas Biossensoriais/instrumentação , Ureia/sangue , Urease/metabolismo , Zeolitas/química , Técnicas Biossensoriais/métodos , Enzimas Imobilizadas/metabolismo , Humanos , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes , Ureia/química , Ureia/metabolismoRESUMO
The aim of this work was to develop an array of biosensors for simultaneous determination of four carbohydrates in solution. Several enzyme systems selective to lactose, maltose, sucrose and glucose were immobilised on the surface of four conductometric transducers and served as bio-recognition elements of the biosensor array. Direct enzyme analysis carried out by the developed biosensors was highly sensitive to the corresponding substrates. The analysis lasted 2 min. The dynamic range of substrate determination extended from 0.001 mM to 1.0-3.0mM, and strongly depended on the enzyme system used. An effect of the solution pH, ionic strength and buffer capacity on the biosensors responses was investigated; the conditions of simultaneous operation of all biosensors were optimised. The data on cross-impact of the substrates of all biosensors were obtained; the biosensor selectivity towards possible interfering carbohydrates was tested. The developed biosensor array showed good signal reproducibility and storage stability. The biosensor array is suited for simultaneous, quick, simple, and selective determination of maltose, lactose, sucrose and glucose.
Assuntos
Técnicas Biossensoriais/instrumentação , Condutometria/métodos , Glucose/análise , Lactose/análise , Maltose/análise , Sacarose/análise , Calibragem , Condutometria/instrumentação , Enzimas Imobilizadas/química , Concentração de Íons de Hidrogênio , Concentração Osmolar , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Soluções , Transdutores , ÁguaRESUMO
A highly sensitive conductometric biosensor for l-arginine determination was developed by exploiting the unique biorecognition capacities of two enzymes of urea cycle - arginase (E.C. 3.5.3.1) and urease (E.C. 3.5.1.5). The enzymes were co-immobilized in a single bioselective membrane on the working sensor, while a lysine rich bovine serum albumin (BSA) membrane was immobilized on the reference sensor, allowing differential measurements. The optimum percentage ratio of arginase and urease within the bioselective membrane was determined when the biosensor sensitivity to l-arginine and urea was optimum. Analytical characteristics of the conductometric biosensor for l-arginine determination were compared for two types of enzyme immobilization (cross-linking with glutaraldehyde (GA) and entrapment in the polymeric membrane). The optimum features in terms of the sensitivity, the linear range, and the detection limit (4.2 µS/mM, 0.01-4mM, and 5.0 × 10(-7)M, respectively) were found for l-arginine biosensor based on enzyme cross-linking with GA. A quantitative determination of l-arginine in the real sample (a drinkable solution "Arginine Veyron") gave a satisfactory result compared to the data provided by the producer (a relative error was 4.6%). The developed biosensor showed high operational and storage stability.
Assuntos
Arginase/química , Arginina/análise , Técnicas Biossensoriais/métodos , Enzimas Imobilizadas/química , Urease/química , Animais , Técnicas Biossensoriais/instrumentação , Bovinos , Condutometria , Reagentes de Ligações Cruzadas/química , Eletrodos , Glutaral/química , Concentração de Íons de Hidrogênio , Limite de Detecção , Membranas Artificiais , Soroalbumina Bovina/químicaRESUMO
A differential pair of planar thin-film interdigitated electrodes, deposited on a ceramic pad, was used as a conductometric transducer. The three-enzyme system (invertase, mutarotase, glucose oxidase), immobilized on the transducer surface, was used as a bioselective element. The ratio between enzymes in the membrane was found experimentally considering the highest biosensor sensitivity to substrate (sucrose) and heavy metal ions. Optimal concentration of sucrose for inhibitory analysis was 1.25 mM and incubation time in the investigated solution amounted to 10-20 min. The developed biosensor demonstrated the best sensitivity toward ions Hg(2+) and Ag(+). A principal possibility of the biosensor reactivation either by EDTA solution after inhibition with silver ions or by cysteine solution after inhibition with mercury ions was shown.