Direct electron transfer reactions of microperoxidase were achieved with the help of semiconductive zinc oxide nanoparticles on a pyrolytic graphite electrode. The enzyme could also exhibit fine ...electrocatalytic activity towards the reduction of hydrogen peroxide. Thereby, a hydrogen peroxide biosensor was constructed based on the electrocatalysis of microperoxidase. Further studies revealed that after irradiating the microperoxidase/zinc oxide nanoparticles co-modified electrode with UV light for 4
h, the catalytic ability of microperoxidase could be greatly promoted, which could be beneficial to developing more sensitive hydrogen peroxide biosensors. As comparison, it was found that the catalytic activity of the enzyme would be depressed if microperoxidase/agarose co-modified electrode was irradiated. We supposed it was the photovoltaic effect of the zinc oxide nanoparticles that improved the catalytic ability of microperoxidase.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
In this paper, we report a novel electrochemical detection approach for platelet-derived growth factor (PDGF) via “sandwich” structure and gold nanoparticles (Au-NPs) mediated amplification ...technique. The “sandwich” structure is fabricated based on the fact that PDGF has two aptamer-binding sites, which makes it possible for one PDGF molecule to connect with two aptamers simultaneously. It is found that this electrochemical system with “sandwich” structure and Au-NPs can significantly amplify the signal of electrochemical probe Ru(NH
3)
5Cl
2+ for PDGF detection, and thus increase the detection sensitivity significantly. As a result, this PDGF detection approach obtains an extraordinarily low detection limit of 1
×
10
−14
M for purified samples, 1
×
10
−12
M for contaminated-ridden samples or undiluted blood serum. This detection approach can also exhibit good stability and excellent specificity.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
► A “signal-on” electrochemical aptasensor simultaneously detects MUC1 and VEGF165 ► Aptasensors not only detect the two markers but also distinguish their co-existence ► The well specificity and ...sensitivity of the detection are also demonstrated.
In this paper, we report a “signal-on” electrochemical aptasensor for simultaneous determination of two tumor markers MUC1 and VEGF165, by using a ferrocene-labeled aptamer-complementary DNA (cDNA) as probe. Since the cDNA immobilized on an electrode surface can hybridize with both MUC1 aptamer and VEGF165 aptamer to form a long double strand with ferrocene far away from the electrode surface, the probe cannot give electrochemical signal. Nevertheless, the presence of the two tumor markers will inhibit the hybridization of cDNA with the aptamers, thus the distance between ferrocene and the electrode is changed, and a “signal-on” electrochemical method to detect two tumor markers is developed. Experimental results show that the electrochemical signal increases with the addition of either tumor markers, but the biggest electrochemical signal can only be obtained when both tumor markers are present. Therefore, the proposed electrochemical aptasensor can not only detect the two markers but also distinguish their co-existence. It may also display high selectivity and sensitivity towards the detection of the tumor markers, so it might have potential clinical application in the future.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
The safety of nucleic acid staining dyes has long been recognized to be a problem. Extensive efforts have been made to search for alternatives to the most popular but toxic staining dye, ethidium ...bromide （EtBr）. However, so far no staining method that can be guaranteed to be suffidently safe has been developed. In this paper, we report a green staining method of DNA in polyacrylamide gel electrophoresis, where in situ synthesis of DNA-templated fluorescent copper nanoclusters （CuNCs） in the gel is achieved to make the DNA bands visible under UV light. Moreover, a comprehensive study of the performance of this staining method has been conducted and the experimental results show that it has favorable sensitivity, stability, and usability. Meanwhile, in our animal experiments, the two reagents （copper sulfate and ascorbic acid） as well as the synthesized CuNCs have been proven to be non-toxic in contact with skin. In addition, all the reagents employed in this work are readily available and low cost, and the procedure is simple to carry out. Therefore, this novel staining method based on the in situ synthesis DNA-templated fluorescent CuNCs has many potential applications.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
A novel strategy for the fabrication of electrochemical aptasensor is proposed in this work, and the strategy has been employed to develop an aptasensor for the assay of adenosine deaminase activity. ...While a well-designed oligonucleotide containing three functional regions (an adenosine aptamer region, a G-quadruplex halves region, and a linker region) is adopted in our strategy as the core element, the enzymatic reaction of adenosine catalyzed by adenosine deaminase plays a key role as well in the regulation of the binding of the G-quadruplex halves with hemin, the electroactive probe, which is to reflect the activity of the enzyme indirectly but accurately. The detection limit of the fabrication biosensor can be lowered to 0.2 U mL−1 of adenosine deaminase, and 1 nM of the inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride is enough to present distinguishable electrochemical response. Moreover, since the electroactive probe is not required to be bound with the oligonucleotide, this strategy may integrate the advantages of both the labeled and label-free strategies.
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IJS, KILJ, NUK, PNG, UL, UM
•As a new kind of activator, lignin can greatly increase α-amylase activity.•Lignin may interact with α-amylase to form 1:1 complex.•Hydrogen bonding plays a key role in the process of the binding.
...This paper reports a new kind of activator of α-amylase, lignin, which can greatly increase α-amylase activity. The promoted ratio of lignin is even much higher than that of chloride ion, the traditional activator of α-amylase. Further experimental results reveal that lignin may interact with α-amylase to form a 1:1 complex with a binding constant of 4.47×105M−1. The binding is spontaneous and lignin/α-amylase complex formation is an exothermal reaction. Hydrogen bonding plays a key role and non-radiation energy transfers from α-amylase to lignin in the binding process. Lignin, combining with α-amylase, conforms to a first-order exponential decay function. The formation of the lignin/α-amylase complex results in the reduction of α-helical content from 57.7% to 53.9%, the increase of the polarity around tryptophan residues, the decrease of the hydrophobicity, and the enlargement of protein granule volume. This work will give a deeper insight into lignin as a kind of dietary fibre, known as an important food functional factor. Furthermore, it also contributes to the exploration of an activator of α-amylase, used in the food industry.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
► Signal-on biosensing is achieved by synchronizing SWV frequency with probe dynamics. ► Simple peptide probe recognizes Aβ 1–42 soluble oligomer specifically. ► Sensor recognizes its target ...quantitatively and works well in biological matrix.
Based on oligopeptide, a novel strategy to fabricate electrochemical biosensor is proposed in this work by fine-tuning the scan pulse frequency of square wave voltammetry (SWV) to synchronize with the surface electron transfer (ET) of the oligopeptide modified on an electrode surface. By using this strategy, the surface ET dynamics of our peptide-based biosensor can show significant difference in the presence and absence of a detection target, thus the proposed strategy has been employed for the assay of amyloid β 1–42 (Aβ 1–42) soluble oligomer, which is among the most neurotoxic species of Aβ peptide. Experimental results reveal that our sensor might be an appropriate candidate for quantitative assay of Aβ 1–42 soluble oligomer. Moreover, the strategy proposed in this work may be extended for the fabrication of more peptide-based biosensors in the future.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
Sensitive and accurate detection of cancer cells plays a crucial role in clinical diagnosis, treatment and prognosis of tumors. In this paper, we report a new electrochemical method for highly ...selective and sensitive detection of cancer cells by using small molecule-linked DNA as probes. The methodology is based on the fact that exonuclease I can catalyze the digestion of folate-linked DNA probes that are immobilized on an electrode surface; however, in the presence of the target cells, such as human breast cancer MCF-7 cells, the probes can be protected from digestion upon the binding with folate receptor that is over-expressed on the cell surface. Consequently, cancer cells can be efficiently detected by monitoring the status of the probe DNA with electrochemical techniques. In this study, the protection to exonuclease I-catalyzed digestion has also been proven by electrochemical studies. Moreover, the proposed method has been proven to linearly detect MCF-7 cells in a wide range from 102–106cellmL−1 with a low detection limit of 67cellmL−1, which can also easily distinguish the folate receptor-negative normal cells, for instance, islet β cells. The reproduction of the detection is also satisfactory, since the relative standard deviations for three independent measurements of different concentration of MCF-7 cells are all within 10%. By replacing the small molecules linked on the DNA probe, other cancer cells can also be detected by making use of this proposed method. Therefore, our cytosensor may have great potential in clinical applications.
•Small molecule-linked DNA has been immobilized on the electrode surface as the probe.•Folate can specially bind to folate receptor over-expressed on the surface of cancer cells.•Cancer cells can be sensitively detected based on the principle of terminal protection.•Proposed biosensor can easily distinguish the folate receptor-negative normal cells.•The method can also be feasible to determinate MCF-7 cells in complex serum samples.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Diagnosis of apoptosis is essential to the early detection of therapy efficiency and the evaluation of disease progression. Caspase-3 is supposed to be closely related to cellular apoptosis. We ...describe here a label-free surface plasmon resonance (SPR) detection of apoptosis based on caspase-3 activity assay through enzyme digestion. An artificial peptide sequence was designed as a substrate of caspase-3 and immobilized on a gold disk through covalent binding. The 4Lys part at the end of the pentadecyl-peptide was designed to form a unique peptide array through electrostatic repulsion. The immobilization of the peptide on the gold surface was carefully characterized by SPR and atomic force microscopy. The catalytic conditions of caspase-3 were optimized with electrochemical impedance spectroscopy. The detection limit of caspase-3 was found at a concentration of 1 pg mL(-1). The activity of caspase-3 in apoptotic cells could also be measured sensitively by the one-step and intuitional SPR response decrease. The fabricated simple and convenient caspase-3 sensor is proposed for application in clinical analysis.
So far, the development of a unique strategy for specific biomolecules activity monitoring and precise drugs release in cancerous cells is still challenging. Here, we designed a ...conformation-switchable smart nanoprobe to monitor telomerase activity and to enable activity-triggered drug release in cancerous cells. The straightforward nanoprobe contained a gold nanoparticle (AuNP) core and a dense layer of 5-carboxyfluorescein (FAM)-labeled hairpin DNA shell. The 3' region of hairpin DNA sequence could function as the telomerase primer to be elongated in the presence of telomerase, resulting in the conformational switch of hairpin DNA. As a result, the FAM fluorescence was activated and the anticancer drug doxorubicin (Dox) molecules which intercalated into the stem region of the hairpin DNA sequence were released into cancerous cells simultaneously. The smart method could specifically distinguish cancerous cells from normal cells based on telomerase activity. It also showed a good performance for monitoring telomerase activity in the cytoplasm by molecular imaging and precise release of Dox triggered by telomerase activity in cancerous cells. These advantages may offer a great potential of this method for monitoring telomerase activity in cancer progression and estimating therapeutic effect.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
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