This work explores Electrochemical Impedance Spectroscopy (EIS) detection for a highly-sensitive quantification of prostate-specific antigen (PSA) in Faradaic (f-EIS) and non-Faradaic modes (nf-EIS). Immobilization of monoclonal antibody specific to PSA (anti-PSA) was performed using 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride and N-hydroxysuccinimide crosslinking agents in order to conjugate carboxylic (-COOH) terminated group of 16-Mercaptoundecanoic acid with amine (-NH3+) on anti-PSA epitope. This approach offers simple and efficient approach to form a strong, covalently bound thiol-gold (SAu) for a reliable SAM layer formation. Studies on the topographic of pristine Au-IDE surface were performed by Scanning Electron Microscopy and Energy Dispersive X-ray Spectroscopy techniques, meanwhile a 3-dimensional optical surface profiler, Atomic Force Microscopy and X-ray Photoelectron Spectroscopy techniques were used to validate the successful functionalization steps on the sensor transducer surface. Detection of PSA in f-EIS mode was carried out by measuring the response in charge transfer resistance (Rct) and impedance change (Z), meanwhile in nf-EIS mode, the changes in device capacitance was monitored. In f-EIS mode, the sensor reveals a logarithmic detection of PSA in a range of 100 ng/ml down to 0.01 ng/ml in Phosphate Buffered Saline with a recorded sensitivity of 2.412 kΩ/log10 ([PSA] ng/ml) and the limit of detection (LOD) down to 0.01 ng/ml. The nf-EIS detection mode yields a logarithmic detection range of 5000 ng/ml down to 0.5 ng/ml, with a sensitivity of 8.570 nF/log10 ([PSA] ng/ml) and an LOD of 0.5 ng/ml. The developed bio-assay yields great device stability, specificity to PSA and repeatability of detection that would pave its way for the future development into portable lab-on-chip bio-sensing system.
A highly sensitive silica-alumina (Si-Al)-modified capacitive non-Faradaic glucose biosensor was introduced to monitor gestational diabetes. Glucose oxidase (GOx) was attached to the Si-Al electrode surface as the probe through amine-modification followed by glutaraldehyde premixed GOx as aldehyde-amine chemistry. This Si-Al (∼50 nm) modified electrode surface has increased the current flow upon binding of GOx with glucose. Capacitance values were increased by increasing the glucose concentrations. A mean capacitance value was plotted and the detection limit was found as 0.03 mg/mL with the regression coefficient value, R² = 0.9782 [y = 0.8391x + 1.338] on the linear range between 0.03 and 1 mg/mL. Further, a biofouling experiment with fructose and galactose did not increase the capacitance, indicating the specific glucose detection. This Si-Al-modified capacitance sensor detects a lower level of glucose presence and helps in monitoring gestational diabetes.
Misfolding and accumulation of the protein alpha synuclein in the brain cells characterize Parkinson's disease (PD). Electrochemical based aluminum interdigitated electrodes (ALIDEs) was fabricated by using conventional photolithography method and modified the surfaces with zinc oxide and gold nanorod by using spin coating method for the analysis of PD protein biomarker. The device surface modified with gold nanorod of 25 nm diameter was used. The bare devices and the surface modified devices were characterized by Scanning Electron Microscope, 3D-Profilometer, Atomic Force Microscope and high-power microscope. The above measurement was also performed to measure the interaction of antibody with aggregated alpha-synuclein for normal, aggregated and aggregated alpha synuclein in human serum and distinguished against 3 control proteins (PARK1, DJ-1 and Factor IX). The detection limit for normal alpha synuclein was 1 f. with the sensitivity of 1 f. on a linear regression (R2 = 0.9759). The detection limit for aggregated alpha synuclein was 10 aM with the sensitivity of 1 aM on a linear regression (R2 = 0.9797). Also, the detection limit of aggregated alpha synuclein in serum was 10 aM with the sensitivity of 1 aM on a linear regression (R2 = 0.9739). These results however indicate that, serum has only minimal amount of alpha synuclein.
Field of generating a surface thin film is emerging broadly in sensing applications to obtain the quick and fast results by forming the high-performance sensors. Incorporation of thin film technologies in sensor development for the better sensing could be a promising way to attain the current requirements. This work predominantly delineates the fabrication of the dielectric sensor using two different sensing materials (Gold and Aluminium). Conventional photolithography was carried out using silicon as a base material and the photo mask of the dielectric sensor was designed by AutoCAD software. The physical characterization of the fabricated sensor was done by Scanning Electron Microscope, Atomic Force Microscope, High Power Microscope and 3D-nano profiler. The electrical characterization was performed using Keithley 6487 picoammeter with a linear sweep voltage of 0 to 2 V at 0.01 V step voltage. By pH scouting, I-V measurements on the bare sensor were carried out, whereby the gold electrodes conducts a least current than aluminium dielectrodes. Comparative analysis with pH scouting reveals that gold electrode is suitable under varied ionic strengths and background electrolytes, whereas aluminium electrodes were affected by the extreme acid (pH 1) and alkali (pH 12) solutions.
A report on the de novo Whole Genome Sequence (WGS) of Marinobacter litoralis SW-45, a moderately salt-tolerant bacterium isolated from the seawater in Malaysia is presented. The strain has a genome size of 3.45 Mb and is capable of producing halophilic lipase, protease and esterase enzymes. Computational prediction of non-coding RNA (ncRNA) genes in M. litoralis SW-45 was performed using standalone software known as the non-coding RNA characterization (nocoRNAc). In addition, a phylogenetic tree showing the evolutionary relationship between the strain and other members of the genus Marinobacter was constructed using 16SrRNA sequence information. A total of 385 ncRNA transcripts, 1124 terminator region, and 2350 Stress Induced Duplex Destabilization sites were predicted. The current WGS shotgun project has provided the relevant genetic information that may be useful for the strain's improvement studies. This manuscript gives the first description of M. litoralis with a complete genome.
A simple, single-masked gold interdigitated triple-microelectrodes biosensor is presented by taking the advantage of an effective self-assembled monolayer (SAM) using an amino-silanization technique for the early detection of a prostate cancer's biomarker, the prostate-specific antigen (PSA). Unlike most interdigitated electrode biosensors, biorecognition happens in between the interdigitated electrodes, which enhances the sensitivity and limit of detection of the sensor. Using the Faradaic mode electrochemical impedance spectroscopy (EIS) technique to quantify the PSA antigen, the developed sensing platform demonstrates a logarithmic detection of PSA ranging from 0.5 ng/ml to 5000 ng/ml, an estimated LOD down to 0.51 ng/ml in the serum, and a good sensor's reproducibility. The sensor's detection range covers the clinical threshold value at 4 ng/ml and the crucial diagnosis 'grey zone' of 4-10 ng/ml of PSA in serum for an accurate cancer diagnosis. The selectivity test revealed an excellent discrimination of other competing proteins, with a recorded detection signals at 5 ng/ml PSA as high as 7-fold increase versus the human serum albumin (HSA) and 8-fold increase versus the human glandular kallikrein 2 (hK2). The stability test showed an acceptable stability of the aptasensor recorded at six (6) days before the detection signal started degrading below 10% of the peak detection value. The developed sensing scheme is proven to exhibit a great potential as a portable prostate cancer biosensor, also as a universal platform for bio-molecular sensing with the versatility to implement nanoparticles and other surface chemistry for various applications.
Herein, a rapid and sensitive current-volt measurement was developed for identifying the IS6110 DNA sequence to diagnose Mycobacterium tuberculosis (TB). An aminated capture probe was immobilized on a 1,1'-carbonyldiimidazole-functionalized interdigitated electrode (IDE) silica substrate, and the target sequence was detected by complementation. It was found that all tested concentrations displayed a higher response in current changes than the control, and the limit of detection was 10 fM. The sensitivity ranged from 1 to 10 fM. The control sequences with single-, triple-mismatch and noncomplementary sequences showed great discrimination. This rapid and easy DNA detection method helps to identify M. tuberculosis for early-stage diagnosis of TB.
Over the past decade, science has experienced a growing rise in nanotechnology with ground-breaking contributions. Through various laborious technologies, nanomaterials with different architectures from 0 D to 3 D have been synthesized. However, the 3 D flower-like organic-inorganic hybrid nanomaterial with the most direct one-pot green synthesis method has attracted widespread attention and instantly become research hotspot since its first allusion in 2012. Mild synthesis procedure, high surface-to-volume ratio, enhanced enzymatic activity and stability are the main factor for its rapid development. However, its lower mechanical strength, difficulties in recovery from the reaction system, lower loading capacity, poor reusability and accessibility of enzymes are fatal, which hinders its wide application in industry. This review first discusses the selection of non-enzymatic biomolecules for the synthesis of hybrid nanoflowers followed by the innovative advancements made in organic-inorganic hybrid nanoflowers to overcome aforementioned issues and to enhance their extensive downstream applications in transduction technologies. Besides, the role of hybrid nanoflower has been successfully utilized in many fields including, water remediation, biocatalyst, pollutant adsorption and decolourization, nanoreactor, biosensing, cellular uptake and others, accompanied with several quantification technologies, such as ELISA, electrochemical, surface plasmon resonance (SPR), colorimetric, and fluorescence were comprehensively reviewed.
The objective of this study was to validate the Malay version of the International Index of Erectile Function (IIEF-15) in patients with lower urinary tract symptoms. Reliability and validity was assessed by using the test-retest while Cronbach's alpha was used to assess internal consistency. Effect size 5was evaluated to assess the sensitivity to change in the pre-transurethral resection of the prostate (TURP) vs post-TURP. Internal consistency was excellent. A high degree of internal consistency was observed for each of the 15 items and five domains (Cronbach's alpha value=0.56 and higher and 0.74 and higher, respectively). Test-retest correlation coefficient for the 15 items and domains scores showed no significant changes. Intraclass correlation coefficient for 15 items and domains were high (ICC=0.59 and above). It can be concluded that the Mal-IIEF-15 is suitable, reliable, valid and sensitive to clinical change in the Malaysian population.
Biosensors operating based on electrical methods are being accelerated toward rapid and efficient detection that improve the performance of the device. Continuous study in nano- and material-sciences has led to the inflection with properties of nanomaterials that fit the trend parallel to the biosensor evolution. Advancements in technology that focuses on nano-hybrid are being used to develop biosensors with better detection strategies. In this sense, titanium dioxide (TiO2) nanomaterials have attracted extensive interest in the construction of electrical biosensors. The formation of TiO2 nano-hybrid as an electrical transducing material has revealed good results with high performance. The modification of the sensing portion with a combination (nano-hybrid form) of nanomaterials has produced excellent sensors in terms of stability, reproducibility, and enhanced sensitivity. This review highlights recent research advancements with functional TiO2 nano-hybrid materials, and their victorious story in the construction of electrical biosensors are discussed. Future research directions with commercialization of these devices and their extensive utilizations are also discussed.
An approach was made to enhance the halophilic lipase secretion by a newly isolated moderate halophilic Marinobacter litoralis SW-45, through the statistical optimization of Plackett-Burman (PB) experimental design and the Face Centered Central Composite Design (FCCCD). Initially, PB statistical design was used to screen the medium components and process parameters, while the One-factor-at-a-time technique was availed to find the optimum level of significant parameters. It was found that MgSO4 · 7H2 O, NaCl, agitation speed, FeSO4 · 7H2 O, yeast extract and KCl positively influence the halophilic lipase production, whereas temperature, carbon source (maltose), inducer (olive oil), inoculum size, and casein-peptone had a negative effect on enzyme production. The optimum level of halophilic lipase production was obtained at 3.0 g L-1 maltose, 1% (v/v) olive oil, 30 °C growth temperature and 4% inoculum volume (v/v). Further optimization by FCCCD was revealed 1.7 folds improvement in the halophilic lipase production from 0.603 U ml-1 to 1.0307 U ml-1 . Functional and biochemical characterizations displayed that the lipase was significantly active and stable in the pH ranges of 7.0-9.5, temperature (30-50 °C), and NaCl concentration (0-21%). The lipase was maximally active at pH 8.0, 12% (w/v) NaCl, and 50 °C temperature. Besides, M. litoralis SW-45 lipase was found to possess the promising industrial potential to be utilized as a biocatalyst for the esterification.
Haemophilia is a blood clotting disorder known as 'Christmas disease' caused when the blood has defect with the clotting factor(s). Bleeding leads various issues, such as chronic pain, arthritis and a serious complication during the surgery. Identifying this disease is mandatory to take the necessary treatment and maintains the normal clotting. It has been proved that the level of factor IX (FIX) is lesser with haemophilia patient and the attempt here is focused to quantify FIX level by interdigitated electrode (IDE) sensor. Single-walled carbon nanotube (SWCNT) was utilized to modify IDE sensing surface. On this surface, dual probing was evaluated with aptamer and antibody to bring the possible advantages. The detection limit with antibody was found to be 1 pM, while aptamer shows 100 fM. Further, a fine-tuning was attempted with sandwich pattern of aptamer-FIX-antibody and antibody-FIX-aptamer and compared. Specific elevation of detection with 10 folds was noticed and displayed the detection at 100 f. in both sandwich patterns. In addition, FIX was detected in the diluted human serum by aptamer-FIX-antibody sandwich, it was found that FIX detected from the dilution factor 1:640. A novel demonstration is with higher discrimination against other clotting factors, XI and VII.
Abdominal aortic aneurysm (AAA) is a serious, life-threatening vascular disease that presents as an enlarged area of the aorta, which is the main artery that carries blood away from the heart. AAA may occur at any location in the aorta, but it is mainly found in the abdominal region. A ruptured AAA causes serious health issues, including death. Traditional imaging techniques, such as computed tomography angiogram, magnetic resonance imaging, and ultrasound sonography, have been used to identify AAAs. Circulating biomarkers have recently become attractive for diagnosing AAAs due to their cost-effectiveness compared to imaging. Insulin-like growth factor 1 (IGF-1), a secreted hormone vital for human atherosclerotic plaque stability, has been found to be an efficient biomarker for AAA identification. In this report, immunosensing was performed by using an InterDigitated electrode (IDE) sensor to detect circulating levels of IGF-1. The detection limit of IGF-1 was found to be 100 fM with this sensor. Moreover, related protein controls (IGF-2 and IGFBP3) were not detected with the same antibody, indicating selective IGF-1 detection. Thus, immunosensing by using an IDE sensor may help to effectively diagnose AAAs and represents a basic platform for further development.
This paper describes the synthesis of graphene-based activated carbon from carbonaceous rice straw fly ash in an electrical furnace and the subsequent potassium hydroxide extraction. The produced graphene has a proper morphological structure; flakes and a rough surface can be observed. The average size of the graphene was defined as up to 2000 nm and clarification was provided by high-resolution microscopes (FESEM and FETEM). Crystallinity was confirmed by surface area electron diffraction. The chemical bonding from the graphene was clearly observed, with -C=C- and O-H stretching at peaks of 1644 cm-1 and 3435 cm-1, respectively. Impurities in the graphene were found using X-ray photoelectron spectroscopy and energy dispersive X-ray spectroscopy. The measured size, according to zeta-potential analysis, was 8722.2 ± 25 nm, and the average polydispersity index was 0.576. The stability of the mass reduction was analyzed by a thermogravimetric at 100 °C, with a final reduction of ~ 11%.
Osteosarcoma is a bone cancer formed by the cells of the bone. Children, young adults, and teens are highly affected by osteosarcoma. Early identification of osteosarcoma is mandatory to improve the treatment and increase the lifespan of the patients. MicroRNA-195 (miR-195) was shown to be a suitable biomarker for osteosarcoma, and the present study describes a sensitive method of miR-195 identification by nuclease digestion in ELISA to detect and quantify the level of miR-195. S1 nuclease catalyzed endo- and exonucleolytic digestion of single-stranded (ss) RNA and DNA on ELISA polystyrene substrate, which helped to identify duplexed miR-195. This method selectively and specifically identified miR-195 without any biofouling interactions and reached the limit of detection at 10 fM within the range from 10 fM to 10 nM. Due to complete digestion of ssDNA, single- and triple-mismatched sequences failed to increase the ELISA signal, indicating specific miRNA detection. Furthermore, human serum spiked with miR-195 did not interfere with the detection, confirming selective identification. This method identified miR-195 at a lower level and will help to diagnose earlier stages of osteosarcoma.
This research comprehends iron-oxide nanoparticle (IONP) production, the apparent metallic nanostructure with unique superparamagnetic properties. Durian-rind-extract was utilized to synthesize IONP and the color of reaction mixture becomes dark brown, indicated the formation of IONPs and the peak was observed at ∼330 nm under UV-visible spectroscopy. The morphological observation under high-resolution microscopies has revealed the spherical shape and the average size (∼10 nm) of IONP. The further support was rendered by EDX-analysis showing apparent iron and oxygen peaks. XRD results displayed the crystalline planes with (110) and (300) planes at 2θ of 35.73° and 63.53°, respectively. XPS-data has clearly demonstrated the presence of Fe2P and O1s peaks. The IONPs were successfully capped by the polyphenol compounds from durian-rind-extract as evidenced by the representative peaks between 1633 and 595 cm-1 from FTIR analysis. The antimicrobial potentials of IONPs were evidenced by the disk-diffusion assay. The obtained results have abundant attention and being actively explored owing to their beneficial applications.
Multidrug resistant bacteria create a challenging situation for society to treat infections. Multidrug resistance (MDR) is the reason for biofilm bacteria to cause chronic infection. Plant-based nanoparticles could be an alternative solution as potential drug candidates against these MDR bacteria, as many plants are well known for their antimicrobial activity against pathogenic microorganisms. Spondias mombin is a traditional plant which has already been used for medicinal purposes as every part of this plant has been proven to have its own medicinal values. In this research, the S. mombin extract was used to synthesise AgNPs. The synthesized AgNPs were characterized and further tested for their antibacterial, reactive oxygen species and cytotoxicity properties. The characterization results showed the synthesized AgNPs to be between 8 to 50 nm with -11.52 of zeta potential value. The existence of the silver element in the AgNPs was confirmed with the peaks obtained in the EDX spectrometry. Significant antibacterial activity was observed against selected biofilm-forming pathogenic bacteria. The cytotoxicity study with A. salina revealed the LC50 of synthesized AgNPs was at 0.81 mg/mL. Based on the ROS quantification, it was suggested that the ROS production, due to the interaction of AgNP with different bacterial cells, causes structural changes of the cell. This proves that the synthesized AgNPs could be an effective drug against multidrug resistant bacteria.
The development of electrical biosensor towards device miniaturization in order to achieve better sensitivity with enhanced electrical signal has certain limitations especially complexity in fabrication process and costs. In this paper, an alternative technique with minor modification in the device structure is presented for signal amplification by implementing ambipolar conduction in the biosensor itself. We demonstrated the field-effect transistor (FET)-based biosensor coupled back-gate for attaining a higher sensitivity with the detection of lower target abundance. To utilize the coupled back-gate as a pre-amplifier, silicon-on-insulator wafer with thicknesses of top-silicon and buried oxide (BOX) layers of 70 nm and 145 nm, respectively were desired. Titanium dioxide (TiO2) nanomaterial was deposited using sol-gel method on the channel which acts as a transducer. Surface functionalization on TiO2 thin film allowed an effective immobilization of anti-cardiac troponin I antibody to interact cardiac troponin I (cTnI). Binding events at each step was validated by X-ray photoelectron spectroscopy (XPS) analysis. Further, electrical characterization (Id-Vd) confirms the potentiality of FET-based biosensor to detect cTnI (represents acute myocardial infarction disease) with the concentration ranges from 10 μg/ml down to 1 fg/ml. The sensitivity of 459.2 nA (g/ml)-1 and lower detection limit of 1 fg/ml were achieved at Vbg = -5 V and Vd = 5 V. The designed device demonstrates its ability to detect lower level of cTnI with pre-amplified electrical signal by back-gate biasing.
In this study, biological deoxygenation of graphene oxide (GO) using an Eclipta prostrata phytoextract was performed via the infusion method. The presence of oxide groups on the surface of graphene and removal of oxides groups by reduction were characterized through morphological and structural analyses. Field emission scanning electron microscopy images revealed that the synthesized GO and rGO were smooth and morphologically sound. Transmission electron microscopy images showed rGO developing lattice fringes with smooth edges and transparent sheets. Atomic force microscopy images showed an increase in the surface roughness of graphite oxide (14.29 nm) compared with that of graphite (1.784 nm) due to the presence of oxide groups after oxidation, and the restoration of surface roughness to 2.051 nm upon reduction. Energy dispersive X-ray analysis indicated a difference in the carbon/oxygen ratio between GO (1.90) and rGO (2.70). Fourier-transform infrared spectroscopy spectrum revealed peak stretches at 1029, 1388, 1578, and 1630 cm-1 for GO, and a decrease in the peak intensity after reduction that confirmed the removal of oxide groups. X-ray photoelectron microscopy also showed a decrease in the intensity of oxygen peak after reduction. In addition, thermogravimetric analysis suggested that rGO was less thermally stable than graphite, graphite oxide, and GO, with rGO decomposing after heating at temperatures ranging from room temperature to 600 °C.
This research is focussed to quantify IGF1 by electroanalytical analysis on InterDigitated electrode surface and characterized by the microscopic observations. For the detection, antibody and aptamer were used to analyze the level of IGF1. The sandwich pattern (aptamer-IGF1-antibody) was designed on the chemically modified IDE surface and reached the limit of detection to 10 fM with 100 folds enhancement in the sensitivity. Different control experiments (absence of IGF1, binding with IGF2 and with non-complementary aptamer) were failed to show the current changes, discriminated the specific detection. A good detection strategy is to complement the currently following imaging systems for AAA.