Different PES nanofibrous membrane parameters effect on the efficacy of immunoassay performance

The contribution of electrospun nanofibrous membranes (e.NFMs) in the biosensing platforms opens up a new prospect for the invention of faster and more sensitive analytical devices. In this paper, we utilized e.NFM of polyethersulfone (PES) as a solid substrate for the protein immobilization through two different approaches: physical and covalent. Scanning electron microscopy (SEM) and Fourier‐transform‐infrared (FTIR) tests were performed to study the effect of plasma treatment on protein immobilization efficacy. Moreover, taking advantage of ELISA technique, the influence of different parameters, namely, nanofibers diameter, membrane thickness, plasma treatment time, an incubation time of ethyl‐3‐(3‐dimethylaminopropyl)‐carbodiimide/N‐hydroxysuccinimide (EDC/NHS), and their ratio on antibody immobilization efficacy through two mentioned approaches, was also assessed.     

Minimization of Interferences in Flow Injection Amperometric Glucose Biosensor Based on Oxidation of Enzymatically‐produced H2O2

One of the major problems in amperometric biosensors based on detection of H₂O₂ produced by enzymatic reaction between oxidase enzymes and substrate is the interference of redox active compounds such as ascorbic acid (AA), dopamine (DA) and uric acid (UA). To minimize these interferences, sodium bismuthate was used for the first time as an insoluble pre‐oxidant in the flow injection (FI) amperometric glucose biosensor at a Glucose oxidase (GOx) immobilized Pt/Pd bimetallic modified pre‐anodized pencil graphite electrode (p.PGE). In this context, these interfering compounds were injected into a flow injection analysis (FIA) system using an injector which was filled with NaBiO₃. Thus, these interferents were converted into their redox inactive oxidized forms before reaching the electrode in the flow cell. While glucose was not influenced by the pre‐oxidant in the injector, the huge oxidation peak currents of the interferents decreased significantly in the biosensor. FI amperometric current time curves showed that the AA, DA and UA were minimized by 96 %, 86 %, and 98 % respectively, in the presence of an equivalent concentration of interferences in a 1.0 mM glucose solution. The proposed FI amperometric glucose biosensor exhibits a wide linear range (0.01–10 mM, R²=0.9994) with a detection limit of 2.4×10^?3 mM. Glucose levels in the artificial serum and two real samples were successfully determined using the fabricated FI amperometric biosensor.     

Promising photoluminescence optical approach for triiodothyronine hormone determination based on smart copper metal–organic framework nanoparticles

A copper metal–organic framework nanoparticles (Cu‐MOF‐NPs) synthesized via simple technique. The prepared Cu‐MOF‐NPs nanoparticles were further characterized using ¹H‐NMR, FE‐SEM/EDX and thermal study (DSC/TGA). The FE‐SEM/EDX, thermal analysis, and NMR spectrum data with the other analysis support the nano‐Cu‐MOF structure and the monomeric unit (n[Cu (AIP)₂(APY)(H₂O)₂].4H₂O) of Cu‐MOF‐NPs. The photoluminescence (PL) studies of triiodothyronine hormone (T3) based on the prepared Cu‐MOF‐NPs investigated. The results revealed that the Cu‐MOF‐NPs might be used as a biosensor in the determination of triiodothyronine hormone (T3) in biological fluids through a significant quenching of the photoluminescence intensity of Cu‐MOF‐NPs at excitation wavelength 492 nm. The calibration plot achieved over the concentration range 0.0–200.0 ng/dL T3 hormone with a correlation coefficient 0.996 and limit of detection (LOD) and quantification (LOQ) 0.198 and 0.60 ng/dL, respectively. The PL spectra are indicating that Cu‐MOF‐NPs has highly selective sensing properties for T3 hormone without interfering with other human many hormones types. This approach considered a promising analytical tool for early diagnosis of the cases of thyroid disease. The mechanism of quenching between the Cu‐MOF‐NPs, and T3 hormone studied. The mechanism was a dynamic type and obtained due to the energy transfer mechanism.     

乳过氧化物酶与伴刀豆球蛋白A共修饰电极的电化学活性研究

用乳过氧化物酶(LPO)和伴刀豆球蛋白A(Con A)共修饰金电极,首次得到了乳过氧化物酶的直接电化学响应,在此基础上研究了乳过氧化物酶对过氧化氢(H2O2)的电催化活性,并研究了一氧化氮(NO)对LPO电催化活性的影响.在Con A的作用下,乳过氧化物酶在循环伏安图中显示1对准可逆的氧化还原峰,表现出薄层电化学行为.在pH 7.4的磷酸缓冲溶液中的表观氧化还原电位为 -190 mV.该共修饰电极对H2O2表现出电催化还原活性,由此构建的传感器对H2O2的检测范围是2.0×10-5 ～4.0×10-3 mol/L.实验发现,微摩尔量级的NO会抑制乳过氧化物酶对H2O2的催化活性.     

硅溶胶-凝胶包埋纳米金和酶的葡萄糖生物传感器

采用溶胶-凝胶技术将金纳米粒子和葡萄糖氧化酶一次性固定于硅溶胶-凝胶的网络结构中,制备了葡萄糖生物电化学传感器并优化了传感器的制备条件。酶电极对葡萄糖具有良好的电化学响应,葡萄糖浓度在0.02~2.0 mmol/L范围内和催化电流呈线性关系,检出限为0.005 mmol/L。酶电极在4 ℃下贮存100 d后对葡萄糖的响应仅下降8%。该酶电极灵敏度高、响应快、稳定性好。     

Modelling Dynamics of Amperometric Biosensors in Batch and Flow Injection Analysis

A mathematical model of amperometric biosensors has been developed. The model is based on non-stationary diffusion equations containing a non-linear term related to Michaelis–Menten kinetic of the enzymatic reaction. Using digital simulation, the influence of the substrate concentration as well as maximal enzymatic rate on the biosensor response was investigated. The digital simulation was carried out using the finite difference technique. The model describes the biosensor action in batch and flow injection regimes.     

Amperometric Detection of Catechol and Catecholamines by Immobilized Laccase from DeniLite

DeniLite laccase immobilized Pt electrode was used for detection of catechol and catecholamines. The enzymatically oxidized substrates were measured amperometrically. The sensitivities are 210, 75, 60 and 45 nA/μM with the upper limits of linear ranges of 58, 40, 55 and 55 μM and the detection limits (S/N=3) of 0.07, 0.2, 0.3 and 0.4 μM for catechol, dopamine (DA), norepinephrine (NEPI) and epinephrine (EPI), respectively. The response time (t_90%) is about 2 seconds for each substrate and the long‐term stability is around 40–50 days with retaining 80% of initial activity. The very fast response and the remarkable long‐term stability are the principal advantages of this sensor. In case of catechol, the pH response of the sensor is mainly determined by enzyme's pH profile, however, in case of catecholamines, both enzyme's pH profile and reversibility of the substrate are operated and the optimal pHs for NEPI and EPI shift towards acidic range compared to that for DA. The presence of ascorbic acid (<50 μM) did not interfere with the measurement.     

Modification of Escherichia coli single-stranded DNA binding protein with gold nanoparticles for electrochemical detection of DNA hybridization

The unique binding event between Escherichia coli single-stranded DNA binding protein (SSB) and single-stranded oligonucleotides conjugated to gold (Au) nanoparticles is utilized for the electrochemical detection of DNA hybridization. SSB was attached onto a self-assembled monolayer (SAM) of single-stranded oligonucleotide modified Au nanoparticle, and the resulting Au-tagged SSB was used as the hybridization label. Changes in the Au oxidation signal was monitored upon binding of Au tagged SSB to probe and hybrid on the electrode surface. The amplified oxidation signal of Au nanoparticles provided a detection limit of 2.17 pM target DNA, which can be applied to genetic diagnosis applications. This work presented here has important implications with regard to combining a biological binding event between a protein and DNA with a solid transducer and metal nanoparticles.     

DNA Interaction with 17α‐Ethinylestradiol Studied Using Electrochemical Biosensors and Biosensing in Solution

The synthetic estrogen 17α‐ethinylestradiol (EE2) is an active component of oral contraceptives. It is considered as an endocrine disrupting compound that, once incorporated into an organism, affects the hormonal balance of animals and humans. In this study we characterized the DNA‐EE2 interaction using an electrochemical biosensor and biosensing in solution phase with the double stranded DNA from salmon sperm and deoxyguanosine monophosphate (dGMP). Differential pulse voltammetry method has been applied based on voltammetric anodic responses of the deoxyguanine (dGuo) and deoxyadenine (dAdo) as well as EE2 in the medium of phosphate buffer solution pH 7.0. Binding of EE2 to the nucleobases leads to a decrease of their anodic signals. Association constant for DNA‐EE2 interaction has been estimated to be about 1.1 ? 10³ L mol^?1 and 1.4 ? 10³ L mol^?1 for dGuo and dAdo responses, respectively. The association is reversible as indicated by decrease of the EE2 response in pure buffer solution due to leaching of EE2 from the surface attached DNA. The DNA‐EE2 association has been confirmed also by UV‐vis spectrometric experiments.     

Voltammetric and Impedimetric Detection of Interaction Between Dacarbazine and Nucleic Acids

Dacarbazine (DTIC) is a chemotherapy drug that is used for the treatment of Hodgkin's lymphoma, malignant melanoma, childhood solid tumors and soft tissue sarcoma. The surface confined interaction between DTIC and nucleic acids was investigated for the first time in this study by using both differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) in combination with disposable pencil graphite electrodes. The oxidation signals of DTIC and guanine were measured before and after interaction process using DPV technique. The interaction of DTIC with nucleic acids; poly[A], poly[G], or double stranded of poly[A]‐poly[T] and poly[G]‐poly[C] was also examined using DPV. Furthermore, EIS technique was utilized for detection of the interaction between DTIC and nucleic acids; ssDNA/dsDNA, poly[A], poly[G], or double stranded poly[A]‐poly[T] and poly[G]‐poly[C].