Recent advancements in biosensor designs toward the detection of intestine cancer miRNA biomarkers.

Cancer diagnosis and treatment have been of broad interest among scientists in the last decades due to the high death rate, widespread occurrence, and recurrence after treatment. The survival rate of cancer patients depends greatly on early detection and appropriate treatments. Therefore developing new technologies applicable to sensitive and specific methods of cancer detection is an inevitable task for cancer researchers. Abnormal miRNA expression is contributed to severe diseases such as cancers and since their expression level and type differ strictly during carcinogenesis and later metastasis and treatments, the improved detection accuracy of these miRNAs would undoubtedly lead to early diagnosis, prognosis, and targeted therapy. Biosensors are accurate and straightforward analytical devices that have had practical applications especially in the last decade. Their domain is still growing through a combination of attractive nanomaterials and amplification methods, leading to innovative biosensing platforms for the efficient detection of miRNAs as diagnostic and prognostic biomarkers. In this review, we will provide the recent developments in biosensors to detect intestine cancer miRNA biomarkers and also discuss the challenges and outcomings of this field.

Development of novel aptasensor for ultra-sensitive detection of myoglobin via electrochemical signal amplification of methylene blue using poly (styrene)-block-poly (acrylic acid) amphiphilic copolymer.

Amplification of electrochemical signal in order to betterment of limit of detection in determination of biomarkers has an important role in early detection of some dangerous diseases such as cancers. For this purpose, in this research, two types of poly (styrene)-block-poly (acrylic acid) amphiphilic copolymer (PS61-b-PAA596 and PS596-b-PAA61) were synthesized by controlled radical polymerization method via reversible addition-fragmentation chain transfer polymerization (RAFT) technique. Chemical structure of block copolymers was confirmed by FT-IR spectroscopy and their surface morphology was assessed by scanning electron microscopy (SEM). Self-assembly of these block copolymers into polymeric vesicles (polymersomes), loading and release efficiency of methylene blue as an electroactive indicator were investigated in DMF and THF solvents. On the basis of our findings PS61-b-PAA596 has better capability for loading and release of MB than PS596-b-PAA61. Then the obtained methylene blue-loaded polymersome successfully used for development of an aptasensor toward determination of trace amounts of myoglobin. The proposed aptasensor showed a wide linear range from 1.0 aM to 1.0 µM with an ultra-low detection limit of 0.73 aM. Applying this amplification strategy, determination of myoglobin in real samples was successfully performed.

Electrochemically activated pencil lead electrode as a sensitive voltammetric sensor to determine gallic acid.

An electrochemical sensor for the determination of some polyphenolic compounds such as Gallic acid (GA) and Galloyl esters was developed using the activated pencil lead electrode (APLE). At first, a study has been made of the optimum conditions for electrochemical activation of the pencil lead electrode. Potentiodynamic and potentiostatic strategies were investigated for activation of the pencil lead electrode and the results show that the potentiodynamic pretreatment gives better performance toward measurement of the polyphenolic compounds. Electrochemical properties of GA were investigated using chronoamperometry and cyclic voltammetry; and some thermodynamic and kinetic variables such as α, n α, and D were calculated. Sensitive differential pulse voltammetry (DPV) technique was applied for the determination of Gallic acid and Galloyl esters in different samples. Enhanced oxidation peak currents of Gallic acid were observed at APLE when compared with non-activated PLE. The calibration graph has two linear ranges of 0.49-24.3 µM and 0.07-0.83 mM, and the obtained limit of detection for S/N = 3 was 0.25 µM. Adsorptive stripping differential pulse voltammetry (AdSDPV) was also conducted to determine Gallic acid and Galloyl esters in sub-micromolar concentration range. Using the AdSDPV method, the limit of detection was improved and calculated to be 5.2 nM. The proposed method was successfully applied for quantification of the total concentration of Gallic acid and Galloyl esters in a variety of real samples such as black and green tea, and mango juice samples, and desirable recovery values indicated the good accuracy of the developed sensor.

The development of an electrochemical DNA biosensor based on quercetin as a new electroactive indicator for DNA hybridization detection.

An electrochemical DNA biosensor was designed for the detection of a specific target DNA after hybridization with a complementary DNA probe immobilized onto a glassy carbon electrode surface. Quercetin was successfully used as a new electroactive indicator for the hybridization detection. Different interactions of quercetin with single-stranded DNA (ss-DNA) and double-stranded DNA (ds-DNA) led to different electrochemical signals, which were recorded as cyclic and differential pulse voltammograms enabling hybridization detection. Various parameters influencing the biosensor performance were evaluated, and optimized conditions were obtained. Also, the detection limit of 83 pM with a relative standard deviation of 4.6% was obtained for the determination of complementary oligonucleotides. Then, the developed biosensor was applied successively for the detection of short sequences of hepatitis C virus (HCV1). The hybridization between the probe (PHCV1) and its complementary sequence (HCV1a) as the target was studied. Some hybridization experiments with noncomplementary oligonucleotides also showed that the suggested DNA sensor responds selectively to the target.

Synthesis of chitosan-grafted poly-acrylic acid (CTS-g-PAA) hydrogel and its potential application in biosensors for signal enhancing and bioanalysis.

Nowadays, hydrogels have been attracted a lot of interest due to their immense potential in different fields such as biomedicine and biotechnology. Biodegradable and biocompatible pH-sensitive chitosan-grafted polyacrylic acid (CTS-g-PAA) hydrogel was synthesized by grafting an acrylic acid monomer onto chitosan at the presence of methylene bisacrylamide as a cross-linking agent and ammonium persulphate as an initiator. FT-IR spectroscopy and scanning electron microscopy (SEM) were used to analyze the properties of the obtained hydrogel. The synthesized hydrogel is suitable for the delivery of many hydrophilic drugs or species. Using a multi-walled carbon nanotube modified-glassy carbon electrode (CNT-GCE), the loading and release conditions of Nile Blue (NB) as an electroactive compound were evaluated utilizing the differential pulse voltammetry (DPV) and cyclic voltammetry (CV). The effect of various parameters on the electrochemical signal of NB was investigated, and the optimal conditions for the efficient performance of hydrogel to delivery of NB were obtained. The electrocatalytic current values show linear dependence to NB concentration in the range of 0.098 - 0.971 µM while the detection limit of this electrochemical platform was 12.3 nM. The unique proposed hydrogel with the electroactive NB has a broad range of possible applications in biosensors for signal enhancement and bioanalysis. Supplementary Information: The online version contains supplementary material available at 10.1007/s10854-021-06939-7.

A novel electrochemical imprinted sensor for ultrasensitive detection of the new psychoactive substance "Mephedrone".

Identification and quantitation of mephedrone as one of the popular new psychoactive substances (NPSs) in biological fluids is important. In this study, a novel electrochemical imprinted sensor was designed for ultrasensitive and selective measurement of mephedrone, based on sol-gel molecular imprinted polymer, polytyramine and functionalized multi-walled carbon nanotube@ gold nanoparticles (f-MWCNT@AuNPs) nanocomposite. The developed electrochemical sensor inherits characteristics of the gold and MWCNTs such as high electrical conductivity, large specific surface area and good biocompatibility. Also, tyramine as an additional monomer was used for fabrication of a strongly adhering film on the surface of the electrode. In the proposed method, the concentration of mephedrone was determined indirectly. The change in the current response of [Fe(CN)6]3-/4- redox probe in the presence and absence of mephedrone molecules was used for indirect measurement of mephedrone molecule in solution. Density functional theory (DFT) was applied to better understanding the interactions between the mephedrone, sol-gel polymer and tyramine from molecular viewpoint. Under the optimized experimental conditions, the calibration curve of the designed sensor was plotted and two dynamic linear ranges from 1 to 10 nM and 10-100 nM with a limit of detection (LOD) as low as 0.8 nM (142 pg ml-1) were obtained. Finally, the fabricated sensor was successfully used to detect the mephedrone in biological samples.

Development of quantum-dot-encapsulated liposome-based optical nanobiosensor for detection of telomerase activity without target amplification.

Reactivation of telomerase, which is observed in more than 85% of all known human tumours, is considered a promising tumour marker for cancer diagnosis. With respect to the biomedical importance of telomerase, we have developed a simple strategy based on liposomal fluorescent signal amplification for highly sensitive optical detection of telomerase activity using liposome-encapsulated cadmium telluride quantum dots. In this strategy, telomerase extracted from A549 cells elongated the biotinylated telomerase substrate primer, which was then immobilized on streptavidine-coated microplate wells. After the hybridization of the telomerase-elongated product with biotinylated capture probe, streptavidin was added to the assembly. In the next step, biotinylated liposome was conjugated with capture probe through streptavidin. Finally, QD-encapsulated liposomes were disrupted by Triton X-100, and the fluorescence intensity of the released QDs was measured to detect telomerase activity. The results showed that the proposed nanobiosensor was able to detect telomerase activity from as few as 10 A549 cells without the enzymatic amplification of telomerase extension products. In short, this method is not only convenient and sensitive, but also has a simple operating protocol and a wide detection range (10-5000 cells). A linear range was observed between 50 and 800 cells with a correlation coefficient of 0.982 and regression equation of y = 0.0444 x + 17.137. The proposed method is economical, more user-friendly, without error-prone PCR, with a wide detection range and simple operating protocol without the requirement for sophisticated equipment. Graphical Abstract Schematic representation of the QD-encapsulated liposome-based strategy to amplify fluorescence signal for optical detection of telomerase activity.

Dopamine-loaded liposome and its application in electrochemical DNA biosensor.

In this study, disruption and lyophilization-rehydration of dopamine-loaded liposome and its application in electrochemical DNA biosensor was investigated. The liposomes containing soyphosphatidylcholine and cholesterol were prepared through thin-layer hydration. First, an investigation was carried out to find an appropriate lysing agent for disruption of prepared liposomes. Differential pulse voltammetry, as a high sensitive electrochemical technique, was used along with a multi-walled carbon nanotubes modified glassy carbon electrode for sensitive electrochemical detection of released dopamine from disrupted liposomes. Various lysing agents were investigated and finally, the disruption of liposomes using methanol was selected without any surfactant, because of its least fouling effect. Then, lyophilization of dopamine-loaded liposomes was carried out using sucrose as cryoprotectant. The electrochemical studies of lyophilized liposomes showed that the remained dopamine in sucrose-protected liposomes was higher than sucrose-free liposomes. Furthermore, sucrose has no interference in electrochemical studies. Then, with the addition of biotin-X-DHPE to liposome formulation, the lyophilized sucrose protected dopamine-loaded biotin-tagged liposomes were prepared and the feasibility of application of them in electrochemical DNA biosensor was investigated as signal enhancer and verified for detection of oligonucleotides.

Design and development of PCR-free highly sensitive electrochemical assay for detection of telomerase activity using Nano-based (liposomal) signal amplification platform.

Telomerase, which has been detected in almost all kinds of cancer tissues, is considered as an important tumor marker for early cancer diagnostics. In the present study, an electrochemical method based on liposomal signal amplification platform is proposed for simple, PCR-free, and highly sensitive detection of human telomerase activity, extracted from A549 cells. In this strategy, telomerase reaction products, which immobilized on streptavidin-coated microplate, hybridized with biotinylated capture probes. Then, dopamine-loaded biotinylated liposomes are attached through streptavidin to biotinylated capture probes. Finally, liposomes are ruptured by methanol and the released-dopamine is subsequently measured using differential pulse voltammetry technique by multi-walled carbon nanotubes modified glassy carbon electrode. Using this strategy, the telomerase activity extracted from 10 cultured cancer cells could be detected. Therefore, this approach affords high sensitivity for telomerase activity detection and it can be regarded as an alternative to telomeric repeat amplification protocol assay, having the advantages of simplicity and less assay time.

Electrosorption and photocatalytic one-stage combined process using a new type of nanosized TiO2/activated charcoal plate electrode.

In the present study, an activated charcoal (AC) plate was prepared by physical activation method. Its surface was coated with TiO2 nanoparticles by electrophoretic deposition (EPD) method. The average crystallite size of TiO2 nanoparticles was determined approximately 28 nm. The nature of prepared electrode was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and Brunauer-Emmett-Teller (BET) surface area measurement before and after immobilization. The electrosorption and photocatalytic one-stage combined process was investigated in degradation of Lanasol Red 5B (LR5B), and the effect of dye concentration, electrolyte concentration, pH, voltage, and contact time was optimized and modeled using response surface methodology (RSM) approach. The dye concentration of 30 mg L(-1), Na2SO4 concentration of 4.38 g L(-1), pH of 4, voltage of 250 mV, and contact time of 120 min were determined as optimum conditions. Decolorization efficiency increased in combined process to 85.65% at optimum conditions compared to 66.03% in TiO2/AC photocatalytic, 20.09% in TiO2/AC electrosorption, and 1.91% in AC photocatalytic processes.

Detection and discrimination of recombinant plasmid encoding hepatitis C virus core/E1 gene based on PNA and double-stranded DNA hybridization.

Development of an electrochemical DNA biosensor for direct detection and discrimination of double-stranded plasmid (ds-Pl) without the need for denaturation of the target plasmid sample using a peptide nucleic acid (PNA) oligomer as the probe is described. This goal was achieved by modification of gold electrode with 6-mercapto-1-hexanol following monolayer self-assembly of cysteine conjugated 20-mer PNA oligomer probe, complementary to the HCV core/E1 region, which binds to ds-Pl and forms PNA/ds-Pl structure. The significant variation in differential pulse voltammetric response of methylene blue on the probe modified electrode upon contacting with complementary double-strand plasmid to form PNA/ds-Pl triplex structure is the principle of target plasmid detection. The results indicated that the reduction peak current was linear with the concentration of complementary strand in the range of 10-300 pg/µl with a detection limit of 9.5 pg/µl.

Direct and rapid electrochemical biosensing of the human interleukin-2 DNA in unpurified polymerase chain reaction (PCR)-amplified real samples.

Electrochemical detection of polymerase chain reaction (PCR)-amplified human interleukin-2 (IL-2) coding DNA sample (399bp size) without any purification and pre-treatment is described. To achieve this goal, a sensor was made by immobilization of a 20-mer oligonucleotide (chIL-2) as the probe on the pencil graphite electrode (PGE). This probe is related to the antisense strand of human interleukin-2 gene. The results showed that the electrode could effectively sense the PCR product of human interleukin-2 DNA by anodic differential pulse voltammetry (ADPV) based on guanine oxidation signal. In order to inhibit PCR components interfering effects and improve biosensing performance, various factors were investigated. We found that the desorption of non-specifically adsorbed components of the unpurified PCR samples from PGE surface is easily achieved by washing of the electrode in washing solution for about 300s. The effectiveness of this procedure was confirmed using purified PCR samples. The selectivity of the sensor was assessed with negative control PCR sample and seven different non-complementary PCR products corresponding to 16S rDNA (bigger than 1500bp) of various bacterial genuses. Diagnostic performance of the biosensor is described and the detection limit is found to be 69pM. The reliability of the electrochemical biosensing results was verified by electrophoresis of the PCR products.

Construction, electrochemically biosensing and discrimination of recombinant plasmid (pEThIL-2) on the basis of interleukine-2 DNA insert.

Construction, electrochemically biosensing and discrimination of recombinant pEThIL-2 plasmid, with 5839bp size, on the basis of interleukine-2 (IL-2) DNA insert are described. Plasmid pEThIL-2 was constructed by PCR amplification of IL-2 encoding DNA and subcloning into pET21a(+) vector using BamHI and SacI sites. The recombinant pEThIL-2 plasmid was detected with a label-free DNA hybridization biosensor using a non-inosine substituted probe. The proposed sensor was made up by immobilization of a 20-mer antisense single strand oligonucleotide (chIL-2) related to the human interleukine-2 gene on the pencil graphite electrode (PGE) as a probe and then the sensing of recombinant pEThIL-2 plasmid was conducted by anodic differential pulse voltammetry (ADPV) based on guanine oxidation signal. Selectivity of the detection was assessed with pET21a(+) non-complementary plasmid, with 5443bp size, lacking IL-2 encoding DNA. Different factors such as electrode activation conditions and washing strategy were tested in order to eliminate the nonspecific adsorption of pET21a(+). We have found that the PGE activation for 300s produces a condition in which desorption of nonspecifically adsorbed plasmids from the electrode surface can be achieved by 300s washing of the electrode in 20mM Tris-HCl buffer solution (pH 7.0) containing 20mM NaCl. Diagnostic performance of the biosensor is described and the detection limit is found to be 10.31pg/microL.