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Exosomes, as next-generation biomarkers, has great potential in tracking cancer progression. They face many detection limitations in cancer diagnosis. Plasmonic biosensors have attracted considerable attention at the forefront of exosome detection, due to their label-free, real-time, and high-sensitivity features. Their advantages in multiplex immunoassays of minimal liquid samples establish the leading position in various diagnostic studies. This review delineates the application principles of plasmonic sensing technologies, highlighting the importance of exosomes-based spectrum and image signals in disease diagnostics. It also introduces advancements in miniaturizing plasmonic biosensing platforms of exosomes, which can facilitate point-of-care testing for future healthcare. Nowadays, inspired by the surge of artificial intelligence (AI) for science and technology, more and more AI algorithms are being adopted to process the exosome spectrum and image data from plasmonic detection. Using representative algorithms of machine learning has become a mainstream trend in plasmonic biosensing research for exosome liquid biopsy. Typically, these algorithms process complex exosome datasets efficiently and establish powerful predictive models for precise diagnosis. This review further discusses critical strategies of AI algorithm selection in exosome-based diagnosis. Particularly, we categorize the AI algorithms into the interpretable and uninterpretable groups for exosome plasmonic detection applications. The interpretable AI enhances the transparency and reliability of diagnosis by elucidating the decision-making process, while the uninterpretable AI provides high diagnostic accuracy with robust data processing by a "black-box" working mode. We believe that AI will continue to promote significant progress of exosome plasmonic detection and mobile healthcare in the near future., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

An electrochemical sensor capable of detecting glutathione (GSH) with high sensitivity and selectivity was developed based on the unique novel electroactive silver-based metal organic framework (Ag-MOF). The Ag-MOF obtained by silver nitrate and 1,3,5-benzoic acid (H 3 BTC) was thoroughly characterized and was modified onto the electrode via facile drop-casting method. The electrochemical response of GSH on the Ag-MOF modified electrode showed a significant reduction in the current signal because the Ag-GSH complex had stronger specific affinity than Ag-H 3 BTC and resulted in the collapse of the Ag-MOF. This sensor demonstrated an extensive linear dynamic range of 0.1 nM-1 µM, along with the low detection limit of 0.018 nM. Additionally, it exhibited good reproducibility, stability, and resistance to interfering compounds. The Ag-MOF modified electrode demonstrated superior performance attributed to its rapid electron transfer rate, outstanding electrochemical redox activity, and specific recognition/competitive reaction. These factors improved both sensitivity and selectivity. The high anti-interference ability allowed for the selective detection of GSH in intricate surroundings. In the real sample testing, the RSD was lower than 3.1% and the recovery was between 98.1 and 103%. This research highlights the potential of Ag-MOFs in developing electrochemical sensors and their promising applications in determining GSH for food screening and early disease diagnosis., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Neurotransmitters (NTs) are endogenous, polar, low-molecular-weight compounds that play multiple pivotal roles in the central nervous system. NTs are involved in communicating information, responding to stress, regulating motor coordination, and allowing interneuronal communication in living organisms. It is essential to determine the distribution of NTs in brain regions to better understand drug dependence and abuse, neurological disorders, psychological disorders, and aging. Monitoring NT levels is also important in diagnosing and avoiding serious illnesses. We here review chromatography-based analytical techniques, including pretreatment methods (e.g., microdialysis and solid-phase microextraction), as well as detection strategies (e.g., MS and electrochemistry), focusing on developments in these techniques over the past 5 years. We then highlight recent advances in electrochemical and fluorescence imaging methods in vivo and the disadvantages and advantages of such technologies, including high spatiotemporal resolution, polymer specificity, and high sensitivity. Finally, we summarize and compare the complementary advantages of chromatography-based analytical techniques and biosensors and discuss trends in the development of NT detection technologies., (© 2022 John Wiley & Sons Ltd.)

Aeromonas salmonicida (A. salmonicida) is an important opportunistic pathogen to aquatic animals that causes severe economic losses to aquaculture, which makes its rapid detection and prevention are critical. In this work, a single-stranded DNA (ssDNA) aptamer (A.s-2) with high specificity to the bacteria was selected by Systematic Evolution of Ligands by Exponential Enrichment (SELEX). The selected aptamer was confirmed with high binding ability and specificity (K d  = 32 ± 8 nM). Furthermore, a novel dual-model colorimetric and ratiometric fluorescent aptasensor was constructed based on the G-quadruplex-modified aptamer and g-C 3 N 4 for sensitive, reliable, and visual detection of the diseased bacteria in fishes. The quantitative detection was achieved in the linear range of 10 3 -10 7  CFU mL -1 with a detection limit of 1.9 × 10 2  CFU mL -1 . Meanwhile, the semi-quantitative detection can also be performed visually through fluorescence or color changes of the solution, which is suitable for the early diagnosis of pathogen infection in grassroots farms. Moreover, the developed aptasensor was successfully applied to detect A. salmonicida infection in zebrafish samples with satisfactory results. This work provides a framework for the rapid detection of pathogens in aquaculture, indicating its great prospects in food safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Chemical biosensing techniques are essential for food analysis and disease diagnosis. Nanomaterials with redox activity show great potential in electrochemical analysis, acting as signal labels or signal amplification unit, which can reflect the targets concentration in foods and biological samples. Here, an ultra-sensitive dual-signal intrinsic self-calibration electrochemical platform for GSH was firstly fabricated based on the novel electroactive nanomaterial of ferrocene-functionalized copper metal-organic framework (Fc-Cu-MOF). Due to the solid-state electrochemical property of cuprous chloride (CuCl), a sharp characteristic peak with an increased signal appears with the coexistence of chloride ions in solution. The stronger specific affinity between Cu + and GSH than that of Cu + and Cl - triggers a "crowding effect" that causes the current signal of CuCl decrease greatly. Meanwhile, the peak current of ferrocene keeps unchanged as an internal reference. Based on the ratio of the peak current variation (ΔI Cu /ΔI Fc ) as the signal output, Fc-Cu-MOF modified electrode showed wider linear range in 0.1 nM -1 μM for GSH with the detection limit as low as 0.025 nM. And the sensor was successfully applied in the determination of GSH with excellent recoveries in various real samples such as food and serum samples, providing good prospect in application of bioanalysis and food screening., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Rod-like graphite carbon nitride@MnO 2 (R-g-C 3 N 5 @MnO 2 ) heterostructure was prepared by in situ self-anchored growth of MnO 2 nanosheet on the surface of R-g-C 3 N 5 . The synthesized R-g-C 3 N 5 @MnO 2 heterostructure as photoactive material exhibited excellent photoelectrochemical (PEC) performance, and the prepared heterostructure-aptamer probe displayed sensitive PEC response to cTnI. Therefore, the PEC method was developed to detect cTnI based on the R-g-C 3 N 5 @MnO 2 heterostructure. It was found that the linear response to cTnI was in the range 0.001-30 ng/mL under optimized conditions, and the detection limit of the proposed sensor was 0.3 pg/mL. The PEC method displays stable photocurrent response up to 8 cycles and exhibited outstanding selectivity and sensitivity. The PEC method was successfully applied to detect cTnI in serum samples. The recoveries of cTnI detection in serums reach 95.5-104%, and the relative standard deviations range from 3.20 to 4.45%., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

An electrochemiluminescence (ECL) sensor provides a sensitive and convenient method for early diagnosis of diseases; however, it is still a challenge to develop simple and sensitive sensing platforms based on efficient ECL signals and luminophore groups. Porphyrin-based metal-organic frameworks (MOFs) show great potential in ECL sensing; however, the mechanism and structure-activity relationship, as well as application, are rarely reported. Herein, hydrothermal reactions obtained porphyrin Zr-MOFs (PCN-222) with different specific surface areas, pore sizes, structures, and surface charge states by tuning the reaction time were developed, which served both as the ECL luminophore, coreaction promoter for S 2 O 8 2- , and a connection in the ECL immunoassay. By progressively controlling the condition of the hydrothermal reaction, PCN-222 with large surface area-abundant micropores can be obtained, which has good conductivity and positively charged surfaces, obtaining excellent ECL performance. The ECL performance and the enhancement mechanism were investigated in detail. Using PCN-222-6h with the best ECL intensity as the immobilization matrix for the aptamer, a highly sensitive and selective assay for thrombin was developed. The decrease of the ECL signal was logarithmically linear with the concentration of thrombin in the range from 50 fg mL -1 to 100 pg mL -1 with a low detection limit of 2.48 fg/mL. This proposed strategy provides a brand new approach for tuning of the structures of MOFs as effective ECL signal probes, thus providing wider possibilities for effective ECL immunoassays in the detection of other biomarkers in diagnosis of diseases.

The abnormal accumulation of copper ions (Cu 2+ ) is considered to be one of the pathological factors of Alzheimer's disease (AD), but the internal relationship between Cu 2+ and AD progression is still not fully clear. In this work, a sensitive and selective near-infrared fluorescent copper ion probe (DDP-Cu) was designed for quantification and visualization of Cu 2+ level in lysates, living cells, living zebrafish and brain tissues of drosophila and mice with AD. By using this probe, we demonstrated that the content of Cu 2+ in the brains of AD mice and drosophila enhanced nearly 3.5-fold and 4-fold than that of normal mice and drosophila, respectively. More importantly, pathogenesis analysis revealed that elevated Cu 2+ led to changes in factors closely associated with AD, such as the increasing of reactive oxygen species(ROS), the aggregation of amyloid-β protein (Aβ) and nerve cell cytotoxicity. These findings could promote the understanding of the roles between Cu 2+ and AD., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A sensitive electrogenerated chemiluminescence (ECL) biosensor for glucose was developed based on in situ growth of TiO 2 nanowires on Ti 3 C 2 MXenes (TiO 2 -Ti 3 C 2 ) as the nanoplatform. Via tuning the alkaline oxidation time, different amount of TiO 2 nanowires can be found on MXenes. An ECL biosensor for glucose was constructed by covalent immobilization of glucose oxidase (GODx) on the glycine functional TiO 2 -Ti 3 C 2 surface, with the ECL signal depending on the in-situ formation of H 2 O 2 via the specifically catalysis of glucose by GODx, resulting in the apparent increase of ECL signal. The TiO 2 -Ti 3 C 2 can also act as the catalyst for the oxidation of H 2 O 2 into O 2 to enhance the ECL of luminol. Based on this strategy, a highly sensitive ECL biosensor for glucose was obtained in wide concentration range of 20 nM-12 mM with a low detection limit of 1.2 nM (S/N = 3). The synergistic effects of large surface area, excellent conductivity, and high catalytic activity of the TiO 2 -Ti 3 C 2 make the sensor highly sensitive toward glucose; the specific enzyme catalysis reaction promises excellent selectivity of the ECL sensor. The proposed biosensor has been employed to detect glucose in human serum, fruits, and sweat samples with excellent performance, providing a universal approach for glucose in various samples, which shows great prospect in clinical diagnostics and wearable sensors., (Copyright © 2021 Elsevier B.V. All rights reserved.)

It remains an unanswered question whether the flux of K + and H + in lysosomes are correlated due to difficulties in simultaneously imaging these two ions. This question is of great value for understanding lysosomal acidification. Herein, we designed DNA quadruplex and triplex based luminescent nanosensors that can, respectively monitor K + and pH in lysosomal lumen. Each sensor contained an upconversion nanoparticle luminophore and a gold nanoparticle quencher, producing green and blue luminescence signals for K + and H + , respectively. The sensors were tested in buffers showing dynamic ranges of 5 to 200 mM K + and pH 5.0 to 8.2. Co-imaging using these two sensors in cells indicated that the influx of H + was accompanied with the efflux of K + , solving this long-standing question of the lysosomal biochemistry., (© 2020 Wiley-VCH GmbH.)

Transition metal carbides/carbonitrides (MXenes), as one of the emerging two-dimensional (2D) materials, have attracted tremendous attention in recent years. Developing MXenes based biosensing platforms is fantastic for the applications of diagnostic and biological research. In this regard, it is pressing need for the synthesis, exfoliation, intercalation, and tuning the surface of MXenes. Compared with the electrochemical implements of MXenes, the optical biosensing applications are in the earlier stage. In this review, the research significance and types of MXenes are introduced followed by the recent progress of synthesis and functionalization of two kinds of titanium carbide (Ti 3 C 2 ) MXenes including nanosheets (NSs) and quantum dots (QDs). In addition, the special properties and applications in optical biosensing platforms including photoluminescence, electrogenerated chemiluminescence (ECL), surface plasmon resonance (SPR), surface-enhanced Raman scattering (SERS), photoelectrochemical (PEC), colorimetric and photothermal sensors are highlighted. Finally, the possible remaining challenges and future trends of Ti 3 C 2 MXenes in optical biosensing platforms are outlined., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Photoelectrochemical (PEC) sensing has emerged as a simple and practical method for the analysis and detection, its separated optical signal and detection electrical signal give it the advantages of reduced background noise and outstanding sensitivity. Here, we synthesized a Ti 3 C 2 /Cu 2 O composite through simple oil bath heating process, whose excellent PEC performance and sensitive photoelectric response to glucose make it a propitious substitution to glucose oxidase. On this basis, we construct a PEC non-enzymatic sensor based on the Ti 3 C 2 /Cu 2 O heterostructure for the detection of glucose. Under the optimal conditions, the photocurrent of Ti 3 C 2 /Cu 2 O is linear with the logarithm value of glucose concentration in the wide range of 0.5 nM to 0.5 mM with a detection limit of 0.17 nM. Furthermore, the successful detection of glucose in standard samples and human serum by the proposed Ti 3 C 2 /Cu 2 O based PEC non-enzymatic sensor demonstrates the application prospect of heterostructure material in PEC sensor, which provides a new thought for the design and construction of PEC non-enzymatic sensing platform., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In recent years, germanium nanoparticles (Ge NPs) have attracted considerable research attention because of its excellent electrical and optical properties. However, it is mainly applied in the field of solar cells, photodetectors and photothermal therapy. Actually, Ge NPs have the advantages of low toxicity and excellent biocompatibility, which means that they have great potential in biosensing applications. However, there are few related reports. In this paper, we discovered that Ge NPs have the intrinsic peroxidase-like catalytic activity and can significantly accelerate the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) by H 2 O 2 for the first time. Compared with natural enzyme (horseradish peroxidase, HRP), Ge NPs maintain higher catalytic stability after treatment at different pH, temperature and ionic strength. Furthermore, a Ge NPs-based colorimetric platform was established for the first time to detect biomolecules with favourable sensitivity and selectivity, such as uric acid, glucose and choline. This work finds the new property of Ge NPs and extends its application in biosensing., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Exosomes, as novel noninvasive biomarkers for disease prediction and diagnosis, have shown fascinating prospects in monitoring cancer-linked public health issues. Herein, a unique Cy3 labeled CD63 aptamer (Cy3-CD63 aptamer)/Ti 3 C 2 MXenes nanocomplex was constructed as a self-standard ratiometric fluorescence resonance energy transfer (FRET) nanoprobe for quantitative detection of exosomes. The Cy3-CD63 aptamer can be selectively adsorbed onto the Ti 3 C 2 MXene nanosheets by hydrogen bond and metal chelate interaction between the aptamer and MXenes, and the fluorescence signal from Cy3-CD63 aptamer was quenched quickly owing to the FRET between the Cy3 and MXenes. The fluorescence of Cy3 greatly recovered after the addition of the exosomes which can specifically combine with the aptamer and release from the surface of Ti 3 C 2 MXenes due to the high affinity between the aptamer and CD63 protein on exosome surface. Meanwhile, the self-fluorescence signal of MXenes in the whole process showed little change, which can be used as a standard reference. Based on the self-standard turn-on FRET biosensing platform the detection limit of exosome was determined as 1.4 × 10 3 particles mL -1 , which was over 1000× lower than that of conventional ELISA method. This fluorescence sensor can also be used for the identification of multiple biomarkers on the exosome surface and different kinds of exosomes, combining with the fluorescent confocal scanning microscope image. The proposed strategy not only provides a universal nanoplatform for exosomes, but also can be extensively expanded to multiple biomarkers detection, which may promise the prospect of MXenes as robust candidates in biological fields.

The selective and sensitive detection of dopamine (DA) is of great significance for the identification of schizophrenia, Huntington's disease, and Parkinson's disease from the perspective of molecular diagnostics. So far, most of DA fluorescence sensors are based on the electron transfer from the fluorescence nanomaterials to DA-quinone. However, the limited electron transfer ability of the DA-quinone affects the level of detection sensitivity of these sensors. In this work, based on the DA can reduce Ag + into AgNPs followed by oxidized to DA-quinone, we developed a novel silicon nanoparticles-based electron transfer fluorescent sensor for the detection of DA. As electron transfer acceptor, the AgNPs and DA-quinone can quench the fluorescence of silicon nanoparticles effectively through the synergistic electron transfer effect. Compared with traditional fluorescence DA sensors, the proposed synergistic electron transfer-based sensor improves the detection sensitivity to a great extent (at least 10-fold improvement). The proposed sensor shows a low detection limit of DA, which is as low as 0.1 nM under the optimal conditions. This sensor has potential applicability for the detection of DA in practical sample. This work has been demonstrated to contribute to a substantial improvement in the sensitivity of the sensors. It also gives new insight into design electron transfer-based sensors., (Copyright © 2018. Published by Elsevier B.V.)

In this paper, the colorimetric sensing of H 2 O 2 related molecules and biothiols based on etching and anti-etching strategy was firstly proposed. Ag/carbon nanocomposite (Ag/C NC) was served as the sensing nanoprobe, which was synthesized via carbon dots (C-dots) as the reductant and stabilizer. The characteristic surface plasmon resonance (SPR) absorbance of Ag nanoparticles (AgNPs) was sensitive to the amount of hydrogen peroxide (H 2 O 2 ). It exhibited strong optical responses to H 2 O 2 with the solution colour changing from yellow to nearly colourless, which is resulted from the etching of Ag by H 2 O 2 . The sensing platform was further extended to detect H 2 O 2 related molecules such as lactate in coupling with the specific catalysis oxidation of L-lactate by lactate oxidase (LOx) and formation of H 2 O 2 . It provides wide linear range for detecting H 2 O 2 in 0.1-80μM and 80-220μM with the detection limit as low as 0.03μM (S/N=3). In the presence of biothiols, the etching from the H 2 O 2 can be hampered. Other biothiols exhibit anti-etching effects well. The strategy works well in detecting of typical biothiols including cysteine (Cys), homocysteine (Hcy) and glutathione (GSH). Thus, a simple colorimetric strategy for sensitive detection of H 2 O 2 and biothiols is proposed. It is believed that the colorimetric sensor based on etching and anti-etching strategy can be applied in other systems in chemical and biosensing areas., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Construction of a new multifunctional chemo/biosensing platform for small biomolecules and tumor markers is of great importance in analytical chemistry. Herein, a novel universal multifunctional nanoplatform for biomolecules and enzyme activity detection was proposed based on fluorescence resonance energy transfer (FRET) between upconversion nanoparticles (UCNPs) and target-inducing enlarged gold nanoparticles (AuNPs). The reductive molecule such as H 2 O 2 can act as the reductant to reduce HAuCl 4 , which will make the Au seeds grow. The enlarged AuNPs can effectively quench the fluorescence of UCNPs owing to the good spectral overlap between the absorption band of the AuNPs and the emission band of the UCNPs. Utilizing the FRET between the UCNPs and enlarged AuNPs, good linear relationship between the fluorescence of UCNPs and the concentration of H 2 O 2 can be found. Based on this strategy, H 2 O 2 related molecules such as l-lactate, glucose, and uric acid can also be quantified. On the basis of UCNPs and PVP/HAuCl 4 , a general strategy for other reductants such as ascorbic acid (AA), dopamine (DA), or enzyme activity can be established. Therefore, the universal multifunctional nanoplatform based on UCNPs and the target-inducing in situ enlarged Au NPs will show its potential as a simple method for the detection of some life related reductive molecules, enzyme substrates, as well as enzyme activity.

Clenbuterol (CLB), a member of β-agonist family, has now been a serious threat to human health due to its illegal usage in the animal feed. In this paper, we designed a fluorescence resonance energy transfer (FRET) system consisting of carbon dots (C-dots) and gold nanoparticles (AuNPs) for recovered fluorescence detecting of CLB. In the presence of CLB, CLB molecules can interact with AuNPs via Au-N bonds, preventing the interaction of C-dots and AuNPs, which induced the recover of the fluorescent intensity. Under the optimal conditions, the limit of detection for CLB was 3 nM, with a wide concentration linear range of 8-200 nM (S/N = 3). Meanwhile, the proposed method was successfully applied to detect CLB in pork samples, illustrating it could be used as a reliable, rapid, and cost-effective method for the determination of CLB residues in pork samples.

The graphitic carbon nitride nanodots (g-C 3 N 4 -dots) were synthesized by a simple electrochemical "tailoring" process from bulk graphitic carbon nitride (g-C 3 N 4 ) under alkaline solution for the first time. Compared with the bulk g-C 3 N 4 , the novel g-C 3 N 4 -dots not only exhibit enhanced fluorescence and excellent dispersion stability in water but also show the reducibility for the reduction of Ag + to AgNPs at 60°C. The biothiols can bound with Ag + through formation of biothiol-Ag + complex to consume the Ag + and act as capping agent to prevent the growth of AgNPs, which cause the decrease of the absorption peak of the AgNPs. Therefore, an optical sensor was developed for the detection of biothiols based on the change of the plasmon resonance absorption peak of the AgNPs. The proposed method exhibits excellent sensitivity and selectivity to biothiols with low detection limit for cysteine (Cys), homocysteine (Hcy), and glutathione (GSH) with 11.5, 16.1, and 15.5nM, respectively. This method also has been successfully applied for the detection of biothiols in human serum with satisfactory results., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this paper, we report a simple and sensitive fluorescent biosensor for the quantitative analysis of silver ions (Ag + ) by using NaYF 4 :Yb 3+ , Tm 3+ upconversion nanoparticles (UCNPs). Ag + could oxidize o-phenylenediamine (OPD) to the oxidized OPD (oxOPD) directly. The fluorescence of UCNPs can be significantly quenched by oxOPD through inner filter effects (IFE). Under the optimized conditions, the Ag + concentration is proportional to the changes of the fluorescence intensity of UCNPs. The proposed method shows high selectivity and Ag + could be quantitatively detected in the range of 0 to 0.5 mM with a low detection limit of 33 nM for Ag + . The selectivity and sensitivity of the detection can also be satisfactory. More importantly, this method has potential in practical application to detect Ag + in real samples without interference.

A new upconversion colorimetric and ratiometric fluorescence detection method for uric acid (UA) has been designed. Yb(3+), Er(3+) and Tm(3+) co-doped NaYF4 nanoparticles (UCNPs) was synthesized. The co-doped NaYF4 nanoparticles, emit upconversion fluorescence with four typical emission peaks centered at 490nm, 557nm, 670nm and 705nm under the 980nm near-infrared (NIR) irradiation. The ZnFe2O4 magnetic nanoparticles (MNPs) possessing excellent peroxidase-like activity was prepared and used to catalyze oxidation the coupling of N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt (TOPS) and 4-amino-antipyrine (4-AAP) in the presence of H2O2 to form purple products (compound 1) which has a characteristic absorption peak located at 550nm. The upconversion fluorescence at 557nm was quenched by the compound 1 while the upconversion emission at 705nm was essentially unchanged, the fluorescence ratio ((I557/I705)0/(I557/I705)) is positively proportional to UA concentration in existence of uricase. More importantly, colorimetric signal can be easily observed and applied to directly distinguish the concentration of UA by the naked eye. Under the optimized conditions, the linear range of colorimetric and ratiometric fluorescence sensing towards UA was 0.01-1mM, the detection limits were as low as 5.79μM and 2.86μM (S/N=3), respectively. The proposed method has been successfully applied to the analysis of UA in human serum. These results indicate that the colorimetric and ratiometric fluorescence dual-readout assay method has great potential for applications in physiological and pathological diagnosis., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A new ratiometric nanosensor is developed for selective and ultrasensitive detection of melamine based on conjugated polyelectrolyte (CPE)-stabilized silver nanoparticles (P1-AgNPs) by perfectly combining the advantages of CPE and AgNPs. P1 featuring a π-delocalized backbone bearing pyridinyl groups can act as an excellent dual-emission fluorescent probe as well as a polymer localizer for AgNPs. In the presence of melamine, the fluorescence intensity at 386nm increases owing to the turn-on of the fluorescence of P1, whereas FL intensity at 488nm decreases due to the melamine-induced aggregation and subsequent aggregation-enhanced emission quenching of P1-AgNPs, therefore leading to the ratiometric fluorescent sensing of analyte. Moreover, analyte-induced aggregation of P1-AgNPs also allows the ratiometric colorimetric measurement of melamine. Under the optimum conditions, this facile ratiometric nanosensor favors the fluorescent and colorimetric determination of melamine in liquid milk products with the detection limit as low as 0.1 and 0.45nM, respectively., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Sudan dyes are banned as food additives due to the carcinogenicity of their metabolites in the human body. Therefore, it is of great significance for sensitive detection of Sudan dyes. This paper reports a novel nanosensor for Sudan dyes detection based on fluorescence (FL) quenching of hexadecyl trimethyl ammonium bromide (CTAB) stabilized upconversion nanoparticles (UCNPs) through the inner filter effect (IFE). In the presence of Sudan I-IV, the fluorescence emission of UCNPs was effectively quenched due to the absorption bands of Sudan I-IV largely covered the emission bands of UCNPs. Under the optimized conditions, the FL was quenched with Sudan concentration over the range of 0.05-40, 0.01-20, 0.01-40 and 0.05-40 μg/mL for Sudan I-IV, respectively. The corresponding limit of detection is 15.1, 2.83, 3.52 and 16.7 ng/mL (at 3σ/slope) respectively. Meanwhile, the nanosensor shows good selectivity, sensitivity and can be successfully applied to detection of Sudan in chili powder samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This paper reports a novel nanosensor for organophosphorus pesticides based on the fluorescence resonance energy transfer (FRET) between NaYF4:Yb,Er upconversion nanoparticles (UCNPs) and gold nanoparticles (AuNPs). The detection mechanism is based on the facts that AuNPs quench the fluorescence of UCNPs and organophosphorus pesticides (OPs) inhibit the activity of acetylcholinesterase (AChE) which catalyzes the hydrolysis of acetylthiocholine (ATC) into thiocholine. Under the optimized conditions, the logarithm of the pesticides concentration was proportional to the inhibition efficiency. The detection limits of parathion-methyl, monocrotophos and dimethoate reached 0.67, 23, and 67 ng/L, respectively. Meanwhile, the biosensor shows good sensitivity, stability, and could be successfully applied to detection of OPs in real food samples, suggesting the biosensor has potentially extensive application clinic diagnoses assays., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A novel label-free fluorescence nanosensor was developed for ultrasensitive detection of protamine and heparin based on fluorescence resonance energy transfer (FRET) between NaYF4:Yb,Er upconversion nanoparticles (UCNPs) and gold nanoparticles (AuNPs). The FRET system was formed by the electrostatic adsorption of AuNPs on UCNPs, and the fluorescence of UCNPs was significantly quenched. When protamine was added to the mixture of UCNPs-AuNPs, the AuNPs interacted with protamine and then desorbed from the surface of UCNPs and aggregated, resulting in the recovery of the fluorescence of UCNPs. On the addition of both protamine and heparin, the FRET system formed owing to the stronger interaction between heparin and protamine than that with AuNPs, leading to a marked fluorescence quenching of UCNPs. The concentrations of protamine and heparin were proportional to the changes of the fluorescence of UCNPs. The linear response range was obtained over the concentration ranges of 0.02 to 1.2μg/ml and 0.002 to 2.0μg/ml with low detection limits of 6.7 and 0.7ng/ml for protamine and heparin, respectively. Simultaneous measurement of protamine and heparin in human serum can be achieved, suggesting that the nanosensor can be used in a complex biological sample matrix., (Copyright © 2015 Elsevier Inc. All rights reserved.)

A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)3(2+) on the rGO electrode interface, affording fast and highly sensitive ECL cytosensing and cell surface glycan evaluation. Combining the multivalent aptamer interface and ALP nanoprobes, the ECL cytosensor showed a detection limit of 38 CCRF-CEM cells per mL in human serum samples, broad dynamic range and excellent selectivity. In addition, the proposed biosensor provided a valuable insight into dynamic profiling of the expression of different glycans on cell surfaces, based on the carbohydrates recognized by lectins applied to the nanoprobes. This biosensor exhibits great promise in clinical diagnosis and drug screening.

We demonstrate a multivalent recognition and highly selective aptamer signal amplification strategy for electrochemical cytosensing and dynamic cell surface N-glycan expression evaluation by the combination of concanavalin A (Con A), a mannose binding protein, as a model, conjugated poly(amidoamine) dendrimer on a chemically reduced graphene oxide (rGO-DEN) interface, and aptamer- and horseradish peroxidase-modified gold nanoparticles (HRP-aptamer-AuNPs) as nanoprobes. In this strategy, the rGO-DEN can not only enhance the electron transfer ability but also provide a multivalent recognition interface for the conjugation of Con A that avoids the weak carbohydrate-protein interaction and dramatically improves the cell capture efficiency and the sensitivity of the biosensor for cell surface glycan. The high-affinity aptamer- and HRP-modified gold nanoparticles provide an ultrasensitive electrochemical probe with excellent specificity. As proof-of-concept, the detection of CCRF-CEM cell (human acute lymphoblastic leukemia) and its surface N-glycan was developed. It has demonstrated that the as-designed biosensor can be used for highly sensitive and selective cell detection and dynamic evaluation of cell surface N-glycan expression. A detection limit as low as 10 cells mL(-1) was obtained with excellent selectivity. Moreover, this strategy was also successfully applied for N-glycan expression inhibitor screening. These results imply that this biosensor has potential in clinical diagnostic and drug screening applications and endows a feasibility tool for insight into the N-glycan function in biological processes and related diseases.

Sensitive, rapid, and simple detection methods for the screening of extensively used organophosphorus pesticides and highly toxic nerve agents are in urgent demand. A novel label-free silicon quantum dots (SiQDs)-based sensor was designed for ultrasensitive detection of pesticides. This sensing strategy involves the reaction of acetylcholine chloride (ACh) with acetylcholinesterase (AChE) to form choline that is in turn catalytically oxidized by choline oxidase (ChOx) to produce betaine and H2O2 which can quench the photoluminescence (PL) of SiQDs. Upon the addition of pesticides, the activity of AChE is inhibited, leading to the decrease of the generated H2O2, and hence the PL of SiQDs increases. By measuring the increase in SiQDs PL, the inhibition efficiency of pesticide to AChE activity was evaluated. It was found that the inhibition efficiency was linearly dependent on the logarithm of the pesticides concentration. Consequently, pesticides, such as carbaryl, parathion, diazinon, and phorate, were determined with the SiQDs PL sensing method. The lowest detectable concentrations for carbaryl, parathion, diazinon, and phorate reached 7.25 × 10(-9), 3.25 × 10(-8), 6.76 × 10(-8), and 1.9 × 10(-7) g/L, respectively, which were much lower than those previously reported. The detecting results of pesticide residues in food samples via this method agree well with those from high-performance liquid chromatography. The simple strategy reported here should be suitable for on-site pesticides detection, especially in combination with other portable platforms.

A new ferrocene benzyne derivative (Fc-SAc) that contained oligo-(phenylene-ethynylene) skeleton, ferrocene and thiolate terminal groups was firstly synthesized. The hydrolysis product of Fc-SAc (Fc-SH) was immobilized onto gold nanoparticles (AuNPs) modified glass carbon electrode (GCE) as sensing element for rutin detection with high sensitivity. The new sensing strategy was proposed by using two Fc-SH modified electrodes: Fc-S/AuNPs/GCE (Electrode1) and Fc-S/AuNPs/graphene-chitosan/GCE (Electrode2). The electrochemical oxidation of rutin on Electrode2 was a diffusion-controlled process, which was different from a mass-controlled process on Electrode1. Under the optimal conditions, the peak currents of the sensors were linearly related to the concentrations of rutin. The linear responses ranges were 0.05-30 μM and 0.04-100 μM with the regression coefficients of 0.998 and 0.997 on Electrode1 and Electrode2, respectively. Electrode2 presented wider linear range, superior high sensitivity, lower detection limit and better stability on determination of rutin., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A novel and ultrasensitive sandwich-type electrochemical aptasensor has been developed for the detection of thrombin, based on dual signal-amplification using HRP and apoferritin. Core/shell Fe(3)O(4)/Au magnetic nanoparticles (AuMNPs) loading aptamer1 (Apt1) was used as recognition elements, and apoferritin dually labeled with Aptamer2 (Apt2) and HRP was used as a detection probe. Sandwich-type complex, Apt1/thrombin/Apt2-apoferritin NPs-HRP was formed by the affinity reactions between AuMNPs-Apt1, thrombin, and Apt2-apoferritin-HRP. The complex was anchored on a screen-printed carbon electrode (SPCE). Differential pulse voltammetry (DPV) was used to monitor the electrode response. The proposed aptasensor yielded a linear current response to thrombin concentrations over a broad range of 0.5-100 pM with a detection limit of 0.07 pM (S/N=3). The detection signal was amplified by using apoferritin and HRP. This nanoparticle-based aptasensor offers a new method for rapid, sensitive, selective, and inexpensive quantification of thrombin, and offers a promising potential in protein detection and disease diagnosis., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Four new conjugated oligo-phenylene-ethynylenes derivatives, N-methyl-4-(4-acetylthiophenylethynyl)-1,8-naphthalimide (1), thioacetic acid S-[4-(4-aminophenyl-ethynyl)phenyl]ester (2), 4-methylthiophenylethynylbenzenamine (3), N-methyl-4-(4-methyl-thiophenyl-ethynyl)-1,8-naphthalimide (4), were synthesized by Sonogashira and Eglinton cross-coupling reactions. The structures of the four compounds were confirmed by (1)HNMR, (13)CNMR, MS and IR and their spectral characteristics were studied by ultraviolet and visible (UV) spectroscopy as well as fluorescence spectroscopy in different medium. It was found that the fluorescence properties of compounds 2 and 3 were notably improved in aqueous solutions in the presence of β-cyclodextrin (β-CD). Spectral analysis supported the suppositions that the fluorescence intensity enhancement was due to the formation of inclusion complex with β-CD. The supramolecular interaction was investigated in detail and the reaction mechanism was provided. A salicylaldehyde determination method in aqueous medium was established based on the supramolecular complex of compound 3. Under the optimum conditions, the supramolecular complex exhibited a dynamic fluorescence response range for salicylaldehyde from 0.6 to 240×10(-6) molL(-1), with a detection limit of 1×10(-8) molL(-1)., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A new ferrocene derivative (1-[(4-amino) phenylethynyl]ferrocene, Fc-NH(2)) was synthesized for the first time. The ferrocene derivative molecule contained the phenylethynyl skeleton, ferrocene and amino groups with excellent electrochemical properties. The graphene/Fc-NH(2) nanocomposite was prepared by mixing graphene solution and Fc-NH(2) solution in one pot and the nanocomposite was utilized to construct a Nafion/graphene/Fc-NH(2) modified glassy carbon electrode (GCE). The ferrocene derivative immobilized on the graphene can enhance the charge-transport ability of the nanocomposite, stabilize the graphene and prevent the leakage of ferrocene. The detection signal of dopamine (DA) was significantly amplified on the Nafion/graphene/Fc-NH(2)/GCE. It was experimentally demonstrated that the signal enhancement results from the synergy amplification effect of graphene and the Fc-NH(2). The oxidation peak currents of DA were linearly related to the concentrations in the range of 5 × 10(-8) to 2 × 10(-4) M with the detection limit of 20 nM in the absence of uric acid (UA) and ascorbic acid (AA). In the presence of 10(-3) M AA and 10(-4) M UA, the linear response range was 1 × 10(-7) to 4 × 10(-4) M, and the detection limit was 50 nM at S/N = 3. Using the proposed Nafion/Fc-NH(2)/graphene/GCE, DA was successfully determined in real samples with the standard addition method.

A highly sensitive and selective electrochemical aptasensor for thrombin was developed. By introducing chitosan-gold nanoparticles and horseradish peroxidase (CS-AuNPs-HRP) conjugates to the sensitive union, the thrombin detection signal was dual amplified. The capture probe was prepared by immobilizing an anti-thrombin aptamer on core-shell Fe(3)O(4)-Au magnetic nanoparticles (AuMNPs) and which was served as magnetic separation material as well. The detection probe was prepared from another anti-thrombin aptamer, horseradish peroxidase (HRP), thiolated CS nanoparticle and gold nanoparticle (CS-AuNPs-HRP-Apt2). In the presence of thrombin, the sandwich structure of AuMNPs-Apt1/thrombin/Apt2-CS-AuNPs-HRP was formed and abundant HRP was captured in it. The resultant conjugates are of magnetic characters and were captured onto the surface of a screen printed carbon electrode (SPCE) to prepare the modified electrode by a magnet located on the outer flank of the SPCE. It was demonstrated that the oxidation of hydroquinone (HQ) with H(2)O(2) was dramatically accelerated by the captured HRP. The electrochemical signal, which correlated to the reduction of BQ (the oxidation product of HQ), was amplified by the catalysis of HRP toward the reaction and the enrichment of HRP on the electrode surface. Under optimized conditions, ultrasensitive and high specific detection for thrombin was realized with the proposed assay strategy. The signal current was linearly correlated to the thrombin concentration in the range of 0.01-10 pM with a detection limit of 5.5 fM (S/N = 3). These results promise extensive applications of this newly proposed signal amplification strategy in protein detection and disease diagnosis.

A novel label-free and sensitive fluorescent aptasensor for the detection of potassium ion (K(+)) was developed based on the horseradish peroxidase-mimicking DNAzyme (HRP-DNAzyme). In this work, we selected a K(+)-stabilized single stranded DNA (ssDNA) with G-rich sequence as the recognition element. In the presence of K(+), the G-rich DNA folded into the G-quadruplex structure, and then hemin can bind to the G-quadruplex structure as a co-factor and form HRP-DNAzyme. 3-(p-Hydroxyphenyl)-propanoic acid (HPPA) can be oxidized by H(2)O(2) into a fluorescent product in the presence of DNAzyme. The fluorescence intensity of the HPPA oxidative product increased with the K(+) concentration. Under the optimal conditions, the fluorescence intensity was linearly related to the logarithm of K(+) concentration in the range of 2.5 μM to 5mM. Other metal ions, such as Na(+), Li(+), NH(4)(+), Mg(2+) and Ca(2+) caused no notable interference on the detection of K(+)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Ferritins are nanoscale globular protein cages encapsulating a ferric core. They widely exist in animals, plants, and microbes, playing indispensable roles in iron homeostasis. Interestingly, our study clearly demonstrates that ferritin has an enzyme-mimic activity derived from its ferric nanocore but not the protein cage. Further study revealed that the mimic-enzyme activity of ferritin is more thermally stable and pH-tolerant compared with horseradish peroxidase. Considering the abundance of ferritin in numerous organisms, this finding may indicate a new role of ferritin in antioxidant and detoxification metabolisms. In addition, as a natural protein-caged nanoparticle with an enzyme-mimic activity, ferritin is readily conjugated with biomolecules to construct nanobiosensors, thus holds promising potential for facile and biocompatible labeling for sensitive and robust bioassays in biomedical applications.

A three-dimensional network PAMAM-Au nanocomposite (3D-PAMAM-Au NC) was prepared by using the first generation polyamidoamine dendrimer (G1 PAMAM) as the dispersant agent. The resultant 3D-PAMAM-Au NC was successfully used as an immobilization matrix for the construction of a reagentless mediator-free horseradish peroxidase (HRP)-based H(2)O(2) biosensor on a multi-walled carbon nanotubes (MWCNTs) modified glassy carbon electrode. With the advantages of the three-dimensional network, the organic-inorganic hybrid materials dramatically facilitate the direct electron transfer of HRP, and good bioelectrocatalytic activity towards H(2)O(2) was demonstrated. Under optimum conditions, the current response of the enzyme modified electrode at -0.30 V was detected. The current response is linearly correlated to H(2)O(2) concentration within the range of 18.00 μM to 20.80 mM with a correlation coefficient of 0.9992 and a sensitivity of 377.78 μA mM(-1) cm(-2). The detection limit was down to 6.72 μM (S/N = 3). Furthermore, the biosensor exhibits some other excellent characteristics, such as high selectivity, short response time, and long-term stability. The 3D-PAMAM-Au NC has proved to be a promising biosensing platform for the construction of mediator-free biosensors, and may find wide potential applications in biosensors, biocatalysis, bioelectronics and biofuel cells.

Successful development of an ultrasensitive and highly specific electrochemical aptasensor for thrombin based on amplification of aptamer-gold nanoparticles-horseradish peroxidase (aptamer-AuNPs-HRP) conjugates was reported. In this electrochemical protocol, aptamer1 (Apt1) was immobilized on core/shell Fe(3)O(4)/Au magnetic nanoparticles (AuMNPs) and served as capture probe. Aptamer2 (Apt2) was dual labeled with AuNPs and HRP and used as detection probe. In the presence of thrombin, the sandwich format of AuMNPs-Apt1/thrombin/Apt2-AuNPs-HRP was fabricated. Remarkable signal amplification was realized by taking the advantage of AuNPs and catalytic reactions of HRP. Other proteins, such as human serum albumin, lysozyme, fibrinogen, and IgG did not show significant interference with the assay for thrombin. Linear response to thrombin concentration in the range of 0.1-60 pM and lower detection limit down to 30 fM (S/N=3) was obtained with the proposed method. This electrochemical aptasensor is simple, rapid (the whole detection period for a thrombin sample is less than 35 min), sensitive and highly specific, it shows promising potential in protein detection and disease diagnosis., (Copyright © 2010 Elsevier B.V. All rights reserved.)

A novel method for DNA point mutation detection using single-base-coded CdS nanoparticle probes is proposed. Target DNA was immobilized on core/shell Fe(3)O(4)/Au magnetic nanoparticles. Single-base-coded CdS nanoparticles, such as guanosine coded CdS (G-CdS), cytidine coded CdS (C-CdS), thymidine coded CdS (T-CdS) and adenosine coded CdS (A-CdS) were used as the probes to identify the mutation sites in DNA strand. The hybridization process of single-base-coded CdS nanoparticle probes with the mutation sites in DNA was monitored using piezoelectric quartz crystal microbalance (QCM). The hybridization of the mutation base in DNA with its complementary base-coded CdS nanoprobes specifically caused significant changes in the resonance frequency of the QCM. Thus the base types of the mutation sites in DNA strand could be identified. The results were further confirmed by fluorescence measurement of CdS. Owing to its operation convenience and cost-effective, this DNA point mutation detection method is expected to hold a great promise in the detection of DNA point mutation and genetic assays.

The processes of adhesion, spreading and proliferation of human mammary cancer cells MCF-7 on two Au electrodes with different surface roughness (R(f) and R(f)=3.2 or 1.1) were monitored and clearly identified with the quartz crystal microbalance (QCM) technique. Analyses of the QCM responses on the resonant frequency shifts (Deltaf(0)) vs. the motional resistance changes (DeltaR(1)) revealed a significant surface-stress effect in the involved courses, in addition to a viscodensity effect and a relatively small mass effect (especially at the smooth electrode). Experiments of fluorescence microscopy, cyclic voltammetry and electrochemical impedance spectroscopy were conducted to investigate the cell population on the electrode vs. the electrode-surface roughness. Simplified equations are deduced to quantitatively evaluate the surface stress, and a novel QCM method for dynamically measuring the surface stress on an electrode in cell-culture course is thus described. It was found that the smoother surface (R(f)=1.1) gave a higher surface stress during cell attachment and less cell population on it than the rougher surface (R(f)=3.2). In addition, real-time QCM monitoring showed on the same electrode the surface stress induced by hepatic normal cells being notably higher than that caused by hepatic cancer cells at cell-attachment stage, suggesting that the surface-stress measurement can exhibit the difference of adhesion-performance between the healthy and ill-behaved cells.

The real-time monitoring of the agglutination process of human hepatic normal cells (L-02) at the quartz crystal microbalance (QCM) gold (Au) electrode was performed. Two lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), induced the cell agglutination, resulting in the different Deltaf(0) and DeltaR(1) responses from those caused by the normal cell attachment and growth. The cell-Con A-cell aggregates had higher affinity for the Au substrate due to the excellent adsorption ability of Con A, which was revealed by increased Deltaf(0) and DeltaR(1) shifts and the obvious mass effect of QCM. In contrast, the lower adsorption ability of cell-WGA-cell aggregates was related to the same characteristic of WGA, presenting the decreased Deltaf(0) and DeltaR(1) responses and the time-extended adhesion phase. Parallel microscopic observation experiments were also carried out and exhibited comparable results. The Deltaf(0) responses during the processes of cell growth and cell agglutination were analyzed using the equations Deltaf(0)=alpha(0)+alpha(1)e(-t/tau(1))+alpha(2)e(-t/tau(2))+alpha(3)e(-t/tau(3)) and Deltaf(0)=alpha(0)+alpha(1)e(-t/tau(1))+alpha(2)e(-t/tau(2)), respectively. Furthermore, the current work proved that the QCM measurement technique based on cell agglutination was useful for discriminating hepatic normal cells (L-02) and hepatic cancer cells (Bel7402).

A protocol of one-pot chemical preoxidation and electropolymerization of monomers (CPEM) in enzyme-containing aqueous suspensions (or solutions) was proposed as a universal strategy for high-activity and high-load immobilization of enzymes to construct amperometric biosensors, which was proven to be effective for the monomer of 1,4-benzenedithiol (BDT), 1,6-hexanedithiol, o-phenylenediamine, o-aminophenol or pyrrole, the preoxidant of K3Fe(CN)6 or p-benzoquinone, and the enzyme of glucose oxidase (GOx) or alkaline phosphatase (AP) to develop GOx-based glucose biosensors or AP-based disodium phenyl phosphate biosensors. As a case examined in detail, a well-dispersed aqueous suspension of the poorly soluble BDT was obtained through its dispersion assisted by ultrasonication and coexisting GOx, which was then subject to chemical preoxidation through adding K3Fe(CN)6, yielding many composites of insoluble BDT oligomers with lots of high-activity enzyme molecules entrapped. Some insoluble composites were then electrochemically codeposited with poly(1,4-benzenedithiol) on an Au electrode, yielding an enzyme film with high-load and high-activity enzyme immobilized. The glucose biosensor prepared here from the CPEM protocol showed much better performance than that from the preoxidant-free conventional electropolymerization (CEP) protocol, with a detection sensitivity increase by a factor of 32 in this case. The GOx-based and AP-based first-generation biosensors developed from the present CPEM protocol all exhibited notably improved performance compared with the analogues from the preoxidant-free CEP protocol. The electrochemical quartz crystal microbalance (EQCM) technique was used to investigate various electrode modification processes. The values of quantity and enzymatic specific activity (ESA) of the immobilized enzymes were evaluated through the EQCM and the conventional UV-vis spectrophotometric method, given that the CPEM protocol notably improved the quantity and the ESA of immobilized enzymes as compared with the preoxidant-free CEP protocol. The proposed CPEM protocol may be interesting in a number of fields, including biosensing, biocatalysis, biofuel cells, bioaffinity chromatography, and biomaterials, and the successful electropolymerization of dithiols in aqueous suspensions (two-phase electropolymerization) may open a new avenue for many monomers that are poorly soluble in neutral aqueous solutions to in situ immobilize biomolecules for bioapplications.

The quartz crystal microbalance method (QCM), in combination with electrochemical impedance spectroscopy (EIS), has been utilized to monitor in situ anti-human IgG adsorption on several Au-based surfaces, bare Au, nanogold/4-aminothiophenol (4AT)/Au, and multi-walled carbon nanotubes (MWCNT)/Au, and succeeding human IgG reactions. Also, the immobilization protocol of anti-human IgG via its glutaraldehyde (GA) cross-linking with self-assembled 4AT on an Au electrode and the subsequent surface immunoreaction were examined. The resonant frequency (f(0)) and the motional resistance (R(1)) of the piezoelectric quartz crystal (PQC) as well as electrochemical impedance parameters were measured and discussed. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) of the ferricyanide/ferrocyanide couple were examined before and after electrode modification, the antibody adsorption and antibody-antigen reactions. We found that the amount for antibody adsorption was the greatest on the colloid Au modified surface, and that at MWCNT ranked the second, while specific bioactivity was almost identical on the four kinds of surfaces. Two parameters simultaneously obtained at the colloid Au modified surface, Deltaf(0) and DeltaC(s) (interfacial capacitance), have been used to estimate the association constant of the immunoreaction.

A flow-through PQC/DNA biosensor system is developed by combining sequential flow polymerase chain reaction (PCR) products denaturing prior to piezoelectric quartz crystal (PQC) detection via hybridization of ssDNA. The PQC/DNA biosensor is fabricated based on complex formation of neutravidin/biotinylated probe in 0.2M NaCl in TE buffer (10mM Tris, 1mM EDTA, pH 7.5). Results show that the coating fabricated provides a desirable quality with satisfactory performance. Its application for Escherichia coli detection under controlled flow at 0.02 mL/min for denaturing PCR products and 10 mL/min for transferring solution between reactors and delivering samples to detector to reduce rehybridization leads to significant improvement in repeatability (R.S.D.<6%, n=5) and sensitivity (DeltaF=34 Hz/1000 E. coli cells) as compared to existing manual method (R.S.D.=19%, n=5 and DeltaF=26 Hz/1000 E. coli cells, respectively). Down to 23 E. coli cells are detected, satisfying the HKEPD requirements for E. coli count in beach water.