MoS2-Polyaniline Based Flexible Electrochemical Biosensor: Toward pH Monitoring in Human Sweat

Wearable pH sensors for sweat analysis have garnered significant scientific attention for the detection of early signs of many physiological diseases. In this study, a MoS₂-polyaniline (PANI) modified screen-printed carbon electrode (SPCE) is fabricated and used as a sweat biosensor. The exfoliated MoS₂ nanosheets are drop casted over an SPCE and are functionalized by a conducting polymer, polyaniline (PANI) via the electropolymerization technique. The as-fabricated biosensor exhibits high super-Nernstian sensitivity of −70.4 ± 1.7 mV pH⁻¹ in the linear range of pH 4 to 8 of 0.1 m standard phosphate buffer solution (PBS), with outstanding reproducibility. The sensor exhibits excellent selectivity against the common sweat ions including Na⁺, Cl⁻, K⁺, and NH₄⁺ with tremendous long-term stability over 180 min from pH 4 to 6. The enhanced active surface area and better electrical conductivity as a consequence of the synergistic effect between MoS₂ and PANI are correlated with the boosted performance of the as-produced biosensor. The feasibility of the sensor is further examined using an artificial sweat specimen and the successful detection confirms the potential of the biosensor for a real-time noninvasive, skin attachable, and flexible wearable pH sensor.     

离子液体在蛋白质电化学中的应用研究

综述了离子液体作为黏合剂、支持电解质和修饰材料这三种功用在氧化还原蛋白质直接电化学中的研究现状,并提出了目前研究存在的问题及此种生物传感器应用前景的展望。     

Encapsulation of urease in double‐layered hydrogels of macroporous poly(2‐hydroxyethyl methacrylate) core and poly(ethylene oxide) outer layer: fabrication and biosensing properties

Double‐layered hydrogels of super‐macroporous poly(2‐hydroxyethyl methacrylate) (PHEMA) cryogel core and poly(ethylene oxide) (PEO) hydrogel outer layer for encapsulation of the enzyme urease were constructed. The enzyme was entrapped into the pores of PHEMA cryogel by soaking and then the core was covered with a PEO layer. The leaking of urease from the core was prevented when the density of the PEO network was increased by incorporation of the crosslinking agent, poly(ethylene glycol) diacrylate. The hybrid system exhibited a nearly constant enzyme activity with time and maintained its structural integrity after several reactions of hydrolysis of urea. The potential of double‐layered hydrogels containing urease for establishment of water pollution with copper was investigated as well. Copyright © 2011 Society of Chemical Industry     

Replacing Mg2+ by Fe2+ for RNA-Cleaving DNAzymes

It has been proposed that Mg²⁺ and Fe²⁺ are very similar in interacting with ribozymes and some protein-based enzymes, but their activities with DNAzymes have yet to be studied. Here, the activity of Fe²⁺ as cofactor for a few RNA-cleaving DNAzymes is investigated. 17E is a well-studied DNAzyme that is active in the presence of many different divalent metal ions; it is highly active with Fe²⁺ with an apparent K_d of 29.7±2.3 μm and a k_obs of 1.12±0.11 min⁻¹ in the presence of 1 mm Fe²⁺ at pH 7.5. Fe²⁺ has 21-fold higher activity than Mg²⁺. Six different DNAzymes are then tested, and only the DNAzymes active with Mg²⁺ (17E, 8–17, and E5) are active with Fe²⁺. Fe²⁺ has 25 and one- to twofold higher activity than Mg²⁺ for the 8–17 and E5 DNAzymes, respectively. In pH>7 buffer and in presence of air, 1 mm Fe²⁺ results in a nonspecific degradation of the DNA strand due to reactive oxygen species (ROS). Cleavage reactions in anoxic environment and antioxidant ascorbate can be used to overcome the effect of oxidation. The findings provide insights for potential DNAzyme catalysis in the early Earth, and they further support the similarity between Mg²⁺ and Fe²⁺ in enzyme catalysis.     

电化学免疫传感器在肿瘤标志物检测中的应用

肿瘤是严重威胁人类健康的疾病之一,降低恶性肿瘤死亡率的主要途径是早期诊断和治疗,肿瘤标志物在肿瘤早期诊断中具有重要的临床应用价值。随着纳米技术的迅猛发展,基于纳米材料构建的电化学传感器可实现对肿瘤标志物的检测,且具有检测灵敏度高、选择性好等优点。本文重点综述了碳纳米材料、贵金属纳米材料、氧化物纳米材料、量子点纳米材料等新型纳米材料电化学免疫传感器的构建原理及其在甲胎蛋白、前列腺抗原、癌胚抗原等肿瘤标志物检测中的应用,分析总结了基于不同纳米材料构建的电化学传感器在各种肿瘤标志物检测中的优缺点,并展望了电化学传感器的发展趋势,提出未来电化学免疫传感器应以微型化、高通量化和商业化为研究重点,并实现对肿瘤标志物的快速、在线、实时检测。     

硅纳米线阵列电极作为细胞色素c传感器的研究

利用化学刻蚀法由p型硅片制备了硅纳米线阵列,经过表面去氧化层处理后,制备了检测蛋白质细胞色素c的电化学传感器.实验表明,硅纳米线阵列电极对细胞色素c有良好的电化学响应,并且在低浓度条件下具备线性响应的特点.根据与未经表面处理的硅纳米线阵列电极的实验结果相对比,提出了细胞色素c所具备的羧基末端与硅纳米线阵列电极表面的Si-H相互作用从而改善传感性能的检测机理.     

Orientation of the Monomeric Porin OmpG in Planar Lipid Bilayers

Outer membrane protein G (OmpG) is a non‐selective porin from Escherichia coli. OmpG is a monomer, which makes it unusual among porins, and suggests that it may be useful in biotechnology. In planar lipid bilayers, individual OmpG pores reconstituted by insertion from detergent exhibit pronounced asymmetry in current‐voltage relationships and voltage‐dependent gating. Here, this asymmetry is used to deduce the orientation of OmpG in the bilayers. We introduced two cysteines into the extracellular loops of OmpG. Cleavage of the disulfide bond formed by these residues significantly increases spontaneous gating of the pore. By adding DTT to one side of the bilayer or the other, we demonstrated that pores showing a quiet trace at negative potentials have a “trans” conformation (extracellular loops on the trans side of the bilayer), while pores showing a quiet trace at positive potentials have a “cis” conformation (extracellular loops on the cis side). With this knowledge, we examined the binding of a cyclodextrin to OmpG. When the cyclodextrin was presented to the extracellular face of the pore, transient multisite interactions were observed. In contrast, when the cyclodextrin was presented to the periplasmic face, a more stable single‐site interaction occurred. Because the cyclodextrin can act as a molecular adapter by binding analytes, this information serves to advance the use of OmpG as a biosensor.     

Observations and analysis of electrokinetically driven particle trapping in planar microelectrode arrays

In situ observations of submicron fluorescent tracers suspended in high ionic strength media sealed in a confined geometry are combined with 3‐D simulations in order to provide a better understanding of the synergism between dielectrophoresis and electrothermal flows that cause rapid particle transport and trapping on the surface of planar quadrupolar microelectrodes. The influence of electrode design on the microfluidic patterns and observed particle collection is examined by employing two different types of microelectrodes in the experiments. The potential use of quadrupolar microelectrodes as means for achieving accelerated sampling and signal amplification in future surface based biosensor devices is illustrated with an experiment involving stable capture of antigen‐coated polystyrene particles on the surface of an antibody‐functionalized microelectrode array.     

A Single Molecular Beacon Probe Is Sufficient for the Analysis of Multiple Nucleic Acid Sequences

Molecular beacon (MB) probes are dual‐labeled hairpin‐shaped oligodeoxyribonucleotides that are extensively used for real‐time detection of specific RNA/DNA analytes. In the MB probe, the loop fragment is complementary to the analyte: therefore, a unique probe is required for the analysis of each new analyte sequence. The conjugation of an oligonucleotide with two dyes and subsequent purification procedures add to the cost of MB probes, thus reducing their application in multiplex formats. Here we demonstrate how one MB probe can be used for the analysis of an arbitrary nucleic acid. The approach takes advantage of two oligonucleotide adaptor strands, each of which contains a fragment complementary to the analyte and a fragment complementary to an MB probe. The presence of the analyte leads to association of MB probe and the two DNA strands in quadripartite complex. The MB probe fluorescently reports the formation of this complex. In this design, the MB does not bind the analyte directly; therefore, the MB sequence is independent of the analyte. In this study one universal MB probe was used to genotype three human polymorphic sites. This approach promises to reduce the cost of multiplex real‐time assays and improve the accuracy of single‐nucleotide polymorphism genotyping.