Reduction of graphene oxide becomes an alternative way to produce a scalable graphene and the resulting nanomaterial namely reduced graphene oxide (rGO) has been utilized in a wide range of potential applications. In this article, the level of green reduction strategies, especially the solution-based reduction methods are overviewed based on recent progression, to get insights towards biomedical applications. The degrees of gaining tips with the solution-based green reduction methods, conditions, complexity and the resulting rGO characteristics have been elucidated comparatively. Moreover, the application of greenly produced rGO in electrochemical biosensors has been elucidated as well as their electrical performance in term of linear range and limit of detections for various healthcare biological analytes. In addition, the characterization scheme for graphene-based materials and the analyses on the reduction especially for the solution-based green reduction methods are outlined for the future endeavours.
The enzyme-linked immunosorbent assay (ELISA) has been widely used for disease surveillance and drug screening due to its relatively higher accuracy and sensitivity. Fine-tuning the ELISA is mandatory to elevate the specific detection of biomolecules at a lower abundance. Towards this end, higher molecular capture on the polystyrene (PS) ELISA surface is crucial for efficient detection, and it could be attained by immobilizing the molecules in the correct orientation. It is highly challenging to immobilize protein molecules in a well-aligned manner on an ELISA surface due to charge variations. We employed a 3-(aminopropyl) triethoxysilane (APTES)- and glutaraldehyde (GLU)-coupled PS surface chemical strategy to demonstrate the high performance with ELISA. A potassium hydroxide treatment followed by an equal ratio of 1% APTES and GLU attachment was found to be optimal, and a longer incubation with GLU favored maximum sensitivity. p24 is a vital early secreting antigen for diagnosing human immunodeficiency virus (HIV), and it has been used for efficient detection with the above chemistry. Three different procedures were followed, and they led to the improved detection of the HIV-p24 antigen at 1 nM, which is a 30-fold higher level compared to a conventional ELISA surface. The surface chemical functionalization shown here also displays a higher specificity with human serum and HIV-TAT. The above approach with the designed surface chemistry could also be recommended for disease diagnosis on other sensing surfaces involving the interaction of the probe and the analyte in heterogeneous test samples.
Silver nanoparticle was synthesized using D-glucosamine chitosan base as green reducing agent at elevated temperature in alkaline pH ranges. The excess of D-glucosamine chitosan base was used as it is both stabilizing and reducing agent at different pHs, regulates the shape and size of the silver nanoparticles. The progressive growth of silver nanoparticles was monitored by UV-Visible spectral studies. A sharp peak at 420 nm indicates the formation of spherical silver nanoparticles. The size and shape of silver nanoparticles were observed from Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) methods. The anisotropically grown nanoparticles were used as probe for Surface Enhanced Raman Studies (SERS) using ATP (4-aminothiophenol) as a model system. The catalytic behavior of silver nanoparticles was exploited for 4-nitrophenol reduction and observed that the reduction reaction follows pseudo first order kinetics with a rate constant 0.65 min. The antibacterial activity of silver nanoparticles was also tested for both gram-positive and -negative microorganisms, in which higher zone of inhibition was observed for gram negative microorganism.
Paracetamol (PAR) is an effective antipyretic and analgesic drug utilized worldwide, safer at therapeutic levels but over-dosing and the chronic usage of PAR results in accumulation of toxic metabolites, which leads to kidney and liver damages. Hence, a simple, rapid, cost-effective, and sensitive analytical technique is needed for the accurate determination of PAR in pharmaceutical and biological samples. Though numerous techniques have been reported for PAR detection, electrochemical methods are being receiving more interest due to their advantages. Moreover, in the past few decades, room temperature ionic liquids (RTILs) have been utilized in electrochemical sensors due to their attractive properties. In this present review, authors gathered research findings available for the determination of PAR using RTIL-based electrochemical sensors and discussed. The advantages and limitations in these systems as well as the future research directions are summarized.
A high-performance interdigitated electrode (IDE) biosensing surface was reported here by utilizing self-assembled silica nanoparticle (SiNP). The modified surface was used to evaluate the complementation of hairpin forming region from Mitoxantrone resistance gene 7 (MXR7; liver cancer-related short gene). The conjugated SiNPs on 3-aminopropyl triethoxysilane functionalization were captured with probe sequence on IDE biosensing surface. The physical and chemically modified surface was used to quantify MXR7 and an increment in the current response upon complementation was noticed. Limit of target DNA detection was calculated (1-10 fM) and this label-free detection is at the comparable level to the fluorescent-based sensing. A linear regression was calculated [y = 0.243x - 0.0773; R² = 0.9336] and the sensitivity was 1 fM on the linear range of 1 fM to 10 pM. With the strong attachment of capture DNA on IDE through SiNP, the surface clearly discriminates the specificity (complementary) versus nonspecificity (complete-, single-, and triple-mismatched sequences). This detection strategy helps to determine liver cancer progression and the similar strategy can be followed for other gene sequence complementation.
This research presents a simple, fast and simultaneous electrochemical quantitative determination of nucleobases, for example guanine (G), adenine (A), and thymine (T) in a beef and chicken livers samples to measure the quality of food products based on hybrids of graphitic carbon nitride/Graphene nanoflakes (g-C3N4/GNF) modified electrode. Graphitic carbon nitride (g-C3N4) made of graphite-like covalent link connects nitrogen, nitride, and carbon atoms in the structural design with improved the electrical properties and low band gap semiconductor. The g-C3N4/GNF nanocomposite was synthesized by the hydrothermal treatment to form a porous g-C3N4 interconnected three dimensional (3D) network of g-C3N4 and GNF. The 3D g-C3N4/GNF/GCE was utilized for the detection of nucleic acid bases with a well resolved oxidation peak for the individual analyte. The electrocatalytic current was established to be a linear range from 0.3 × 10-7 to 6.6 × 10-6, 0.3 × 10-7 to 7.3 × 10-6, and 5.3 × 10-6 to 63.3 × 10-4 M for G, A, and T with a detection limit of 4.7, 3.5 and 55 nM, respectively. The diffusion co-efficient and the kinetic parameters were derived from the chronoamperometry technique. The proposed sensing strategy has been effectively used for the application in real sample analysis and observed that the electrode free from the surface fouling.
This work focused on the detection of cortisol on an interdigitated electrode sensor surface using an anti-cortisol antibody. To improve immobilization, antibodies were conjugated with silver nanoparticles and attached to the surface of the sensor. Cortisol interacted in a dose-dependent manner on the antibody-immobilized sensor surface, and current changes were observed. Linear regression analysis was performed by a 3σ calculation, and the limit of detection fell into the range of 0.01 and 0.1 ng/mL. The sensitivity of cortisol was calculated to be 0.01 ng/mL and the sensor discriminated against other hormones, namely norepinephrine and progesterone, with higher selectivity for cortisol. This result represented the selective detection of cortisol with high performance, which can help to determine anxiety disorders.
Tuberculosis (TB) is a highly contagious life-threatening disease caused by the bacterial pathogen Mycobacterium tuberculosis. ESAT-6, an abundant early secretory antigenic target protein by M. tuberculosis, found to play a vital role in virulence. Developing a friendly method for the detection of ESAT-6 at the lower concentration facilitates to treat TB at an earlier stage and helps to control the spreading of disease. Herein, a new single-step approach was designed and was done by pre-mixing ESAT-6 and antibody before being added to the gold nanoparticle (GNP) followed by the salt-induced aggregation. We could attain the detection limit of 1.25 pM, showing the aggregation of GNP and the red spectral shift. Further, a higher specificity was demonstrated with the lack of electrostatic biofouling by ESAT-6 on GNP and retained the dispersed GNP in the presence of 10-kDa culture filtrate protein from M. tuberculosis. The required precise antibody concentration for this assay was found to be 60 nM. The increment in the antibody concentration from 75 nM drastically diminishes the sensitivity to ~ 680-fold, due to the crowding effect. With this assay, we attested the suitability of colorimetric assay for efficiently detecting the smaller-sized protein.
Graphene is a 2-dimensional nanomaterial with an atomic thickness has attracted a strong scientific interest owing to their remarkable optical, electronic, thermal, mechanical and electrochemical properties. Graphene-based materials particularly graphene oxide and reduced graphene oxide are widely utilized in various applications ranging from food industry, environmental monitoring and biomedical fields as well as in the development of various types of biosensing devices. The richness in oxygen functional groups in the materials serves as a catalysis for the development of biosensors/electrochemical biosensors which promotes for an attachment of biological recognition elements, surface functionalization and compatible with micro- and nano- bio-environment. In this review, the graphene-based materials application in electrochemical biosensors based on recent advancement (e.g; the surface modification and analytical performances) and the utilization of such biosensors to monitor the noncommunicable diseases are presented. The detection performances of the graphene-based electrochemical biosensors are in the range of ng/mL and have reached up to fg/mL in detecting the targets of NCDs with higher selectivity, sensitivity and stability with good reproducibility attributes. We have discussed the advances while addressing the very specific biomarkers for the NCDs detection. Challenges and possible future research directions for the NCDs detection based on graphene nanocomposite with other 2D nanomaterials are outlined.
Human Papillomavirus (HPV) is a standout amongst the most commonly reported over 100 types, among them genotypes 16, 18, 31 and 45 are the high-risk HPV. Herein, we designed the oligonucleotide probe for the detection of predominant HPV type 16 for the sensing applications. Conserved amino acid sequences within E6 region of the open reading frame in the HPV genome was used as the basis to design oligonucleotide probe to detect cervical cancer. Analyses of E6 amino acid sequences from the high-risk HPVs were done to check the percentage of similarity and consensus regions that cause different cancers, including cervical cancer. Basic local alignment search tools (BLAST) have given extra statistical parameters, for example, desire values (E-values) and score bits. The probe, 'GGG GTC GGT GGA CCG GTC GAT GTA' was designed with 66.7% GC content. This oligonucleotide probe is designed with the length of 24 mer, GC percent is between 40 and 70, and the melting point (Tm) is above 50°C. The probe needed an acceptable length between 22 and 31 mer. The choice of region is identified here can be used as a probe, has implications for HPV detection techniques in biosensor especially for clinical determination of cervical cancer.
This study presents a novel sulfur-doped graphitic carbon nitride (S@g-C3 N4 ) with a wider potential range as electrocatalyst for electrochemical sensor application. The S@g-C3 N4 nanosheets were successfully prepared with a ball milling method by mixing appropriate molar concentration required precursors. The as-synthesized heteroatom-doped graphitic carbon nitride is characterized by spectroscopic techniques including PL, DRS-UV, FT-IR, and Brunauer-Emmett-Teller equation. The morphological features were studied by FE-SEM and HR-TEM analysis. Chit-S@g-C3 N4 -modified glassy carbon electrode (GCE) was employed for the electrochemical detection of omeprazole (OMZ) use in drug formulations. We have noted an oxidation peak current response at a potential of +0.8 V versus Ag/AgCl in PBS medium (0.1 M, pH 7.0). Differential pulse voltammetry amperometry experimental method can be used to measure the concentration of OMZ for quantitative studies in known samples. Under the optimized experimental condition, the calibration plot was constructed by plotting the peak currents versus OMZ in the linear ranges from 6.0 × 10-7 to 26 × 10-5 M. The linear regression equation is estimated to be Ip (μA) = 0.9518 (C/μM) + 0.3340 with a good correlation coefficient of 0.9996. The lower determination limit was found to be 20 nM and the current sensitivity was calculated (31.722 μA μM-1 cm-2 ). The developed sensor was utilized successfully to determine the OMZ concentration in drug formulations and biological fluids. These results revealed that the Chit-S@g-C3 N4 -modified GCE showed excellent electroanalytical performance for the detection of OMZ at a low LOD, wider linear range, high sensitivity, good reproducibility, long-term storage stability, and selectivity with an acceptable relative standard deviation value.
We demonstrate a green chemistry approach to synthesize narrow-sized zerovalent iron (nZVI) nanoparticles using Artocarpus heterophyllus Lam. leaf extract as reducing and capping agent. The produced nZVI was characterized by various instrumental methods including ultraviolet-visible spectra, transmission electron microscopy, vibrating sample magnetometer (VSM), X-ray diffraction, and Fourier transform infrared spectroscopy. Based on the electron microscopy observations, the particle size was estimated to be ∼30 nm. In VSM, the saturation point of magnetization was observed to be 0.6 emu g-1 under a magnetic field of 0 ± 30 kOe. The synthesized nZVI was amorphous in nature as per the XRD results. The catalytic activity of the nZVI was employed for the catalytic reduction of 4-nitrophenol (4-NP) and decoloration of textile dyes such as methylene blue, methyl orange, and malachite green, respectively. The proposed nZVI synthesis method exhibited better catalytic performance toward reduction of 4-NP and degradation of dyes within 4 min for 0.1 mg of catalyst. Moreover, the synthesized catalyst nZVI can be recoverable and reutilized in many cycles without loss of its significant catalytic activity. The synthesized nZVI could be a promising material to treat industrial wastewater via profitable, sustainable, and ecofriendly approaches.
The current world condition is dire due to epidemics and pandemics as a result of novel viruses, such as influenza and the coronavirus, causing acute respiratory syndrome. To overcome these critical situations, the current research seeks to generate a common surveillance system with the assistance of a controlled Internet of Things operated under a Gaussian noise channel. To create the model system, a study with an analysis of H1N1 influenza virus determination on an interdigitated electrode (IDE) sensor was validated by current-volt measurements. The preliminary data were generated using hemagglutinin as the target against gold-conjugated aptamer/antibody as the probe, with the transmission pattern showing consistency with the Gaussian noise channel algorithm. A good fit with the algorithmic values was found, displaying a similar pattern to that output from the IDE, indicating reliability. This study can be a model for the surveillance of varied pathogens, including the emergence and reemergence of novel strains.
Enzyme Linked Immunosorbent Assay (ELISA) is the gold standard assay for detecting and identifying biomolecules using antibodies as the probe. Improving ELISA is crucial for detecting disease-causing agents and facilitating diagnosis at the early stages of disease. Biotinylated antibody and streptavidin-conjugated horse radish peroxide (streptavidin-HRP) often are used with ELISA to enhance the detection of various kinds of targets. In the present study, we used a competition-based strategy in which we pre-mixed free biotin with streptavidin-HRP to generate high-performance system, as free biotin occupies some of the biotin binding sites on streptavidin, thereby providing more chances for streptavidin-HRP to bind with biotinylated antibody. ESAT-6, which is a protein secreted early during tuberculosis infection, was used as the model target. We found that 8 fM of free biotin mixed with streptavidin-HRP anchored the higher detection level of ESAT-6 by four-fold compared with detection without free biotin (only streptavidin-HRP), and the limit of detection of the new method was 250 pM. These results suggest that biotin-streptavidin competition can be used to improve the diagnosis of analytes in other types of sensors.
Abdominal aortic aneurysm (AAA), a medical complication, occurs when the aortic area becomes swollen and very large. It is mandatory to identify AAA to avoid the breakdown of aneurysms. C-reactive protein (CRP) has been recognized as one of the biomarkers for identifying AAA due to the possibility of CRP produced in vascular tissue, which contributes to the formation of an aneurysm, and it is elevated in patients with a ruptured AAA. This research work was designed to develop an immunosensor on a multiwalled carbon nanotube (MWCNT)-modified surface to quantify the CRP level. Anti-CRP specificity was constructed on the MWCNT surface through a silane linker to interact with CRP. The detection limit of CRP was calculated as 100 pM with an R2 (determination coefficient) value of 0.9855 (y = 2.3446x - 1.9922) on a linear regression graph. The dose-dependent linear pattern was registered from 200 to 3000 pM and attained the saturation level during binding at 3000 pM. Furthermore, serum-spiked CRP showed a clear increase in the current response, proving the specific recognition of CRP in biological samples. This designed biosensor identifies CRP at a lower level and can help diagnose AAA.
Cancer is the uncontrollable abnormal division of cell growth, caused due to the varied reasons. Cancer can be expressed in any part of the body, and it is one of the death-causing diseases. Human reproductive organs are commonly damaged by cancer. In particular, the women reproductive system is affected by various cancers including ovarian, cervical, endometrial, vaginal, fallopian tube, and vulvar cancers. Identifying these cancers at earlier stages prevents the damage to the organs. Aptamer is the potential probe that can identify these cancers. Aptamer is an artificial antibody selected from the randomized library of molecules and has a high binding affinity to the target biomarker. Targeting cancers in the reproductive organs using aptamers showed an excellent efficiency of detection compared to other probes. Different aptamers have been generated against the gynaecological cancer biomarkers, which include HE4, CA125, VEGF, OCCA (for ovarian cancer), EGFR, FGFR1, K-ras (for endometrial cancer), HPV E-16, HPV E-7, HPV E-6, tyrosine, and kinase (for cervical cancer), which help to identify the cancers in woman reproductive organs. In this overview, the biomarkers for gynecologic cancers and the relevant diagnosing systems generated using the specific aptamers are discussed. Furthermore, the therapeutic applications of aptamer with gynaecological cancers are narrated.
Sepsis is an extreme condition involving a physical response to severe microbial infection and causes fatal and life-threatening issues. Sepsis generates during the chemicals release with the immune system into the bloodstream for fighting against an infection, which causes the inflammation and leads to the medical emergency. A complexed longitudinal zeolite and iron oxide nanocomposite was extracted from coal mine fly ash and utilized to improve the surface characteristics of the capacitance biosensor to identify sepsis attacks. Anti-interleukin-3 (anti-IL-3) antibody was attached to the zeolite- and iron oxide-complexed capacitance electrode surface through an amine linker to interact with the sepsis biomarker IL-3. The morphological and chemical components of the nanocomplex were investigated by FESEM, FETEM, and EDX analyses. At approximately 30 nm, the longitudinal zeolite and iron oxide nanocomposite aided in attaining the limit of IL-3 detection of 3 pg/mL on the linear curve, with a regression coefficient (R2) of 0.9673 [y = 1.638x - 1.1847]. A lower detection limit was achieved in the dose-dependent range (3-100 pg/mL) due to the higher amount of antibody immobilization on the sensing surface due to the nanomaterials and the improved surface current. Furthermore, control experiments with relevant biomolecules did not show capacitance changes, and spiked IL-3 in human serum increased capacitance, indicating the specific and selective detection of IL-3. This study identifies and quantifies IL-3 via potentially useful methods and helps in diagnosing sepsis attack.
The primary reasons for myocardial infarction (MI) are pericarditis, arrhythmia, and heart failure, causing predominant deaths worldwide. Patients need a potential diagnostic system and treatment before cardiomyocyte damage. Cardiac biomarkers are released from myocytes immediately after a heart attack. Troponin is an efficient biomarker released from dead cells within a few hours. Aptamers are artificial antibodies used effectively in the biosensor field for biomarker detection. Along with aptamers, the application of nanomaterials is also expected to enhance the detection limits of biosensors. In this investigation, selected aptamers against cardiac troponin I (cTnI) were conjugated with gold nanoparticles (GNPs) to diagnose MI and compared with an aptamer-only control group on an interdigitated electrode surface. Based on electroanalysis, cTnI was detected at concentrations as low as 1 fM, and the detection limit improved to 100 aM when the aptamer was conjugated with GNP. In addition, aptamer-GNP conjugates increased the current level at the tested concentrations of cTnI. Control experiments with noncomplementary aptamers and relevant proteins did not result in notable changes in the current, demonstrating the selective detection of cTnI.
This study correlated and quantified the expression of microRNA-155 with breast cancer to determine breast cancer progression. The target microRNA-155 sequence was identified by complementation on a capture-probe sequence-immobilized interdigitated dual electrode surface. The sensitivity was found to be 1 fM, and the limit of detection fell between 1 and 10 fM. The specific sequence selectivity with single mismatches, triple mismatches, and noncomplementary bases failed to complement the capture-probe sequence. The obtained results demonstrate the selective determination of the microRNA-155 sequence and can help to diagnose breast cancer.