Technological advancements in bio-recognition using liquid crystals: Techniques, applications, and performance.

The application of liquid crystal (LC) materials has undergone a modern-day renaissance from its classical use in electronics industry as display devices to new-fangled techniques for optically detecting biological and chemical analytes. This review article deals with the emergence of LC materials as invaluable material for their use as label-free sensing elements in the development of optical, electro-optical and electrochemical biosensors. The property of LC molecules to change their orientation on perturbation by any external stimuli or on interaction with bioanalytes or chemical species has been utilized by many researches for the fabrication of high sensitive LC-biosensors. In this review article we categorized LC-biosensor based on biomolecular reaction mechanism viz. enzymatic, nucleotides and immunoreaction in conjunction with operating principle at different LC interface namely LC-solid, LC-aqueous and LC-droplets. Based on bimolecular reaction mechanism, the application of LC has been delineated with recent progress made in designing of LC-interface for the detection of bio and chemical analytes of proteins, virus, bacteria, clinically relevant compounds, heavy metal ions and environmental pollutants. The review briefly describes the experimental set-ups, sensitivity, specificity, limit of detection and linear range of various viable and conspicuous LC-based biosensor platforms with associated advantages and disadvantages therein.

Optical absorption investigations for efficient crystal violet dye removal from wastewater via carbon nanotubes:Montmorillonite-based nanocomposite.

The current study reports a facile method to fabricate functionalized multi-walled carbon nanotubes and montmorillonite clay mineral-based nanocomposite matrix and its detailed characterization using spectroscopic and morphological techniques. The nanocomposites have been studied for their potential applications in the treatment of contaminated water using batch adsorption studies. The investigations conducted using optical absorption spectroscopic measurements for the adsorption process indicate that the nanocomposite matrix can effectively remove almost 98% of the dye from aqueous solution. The nanocomposites have showed fast and strong adsorption behaviour for the dye with the maximum adsorption capacity (qm ) of ~467.3 mg g-1 in 25 min. The experimental data at equilibrium were also correlated with the theoretical adsorption isotherm and kinetic models. The results demonstrate that the experimental data fits well to the Freundlich adsorption isotherm model and conforms to second-order kinetics. Furthermore, the nanocomposite exhibits good recyclability without any marked decrease in the adsorption performance even after five adsorption cycles of usage which indicates its potential application as reusable adsorbent for the efficient removal of hazardous dyes from contaminated water.

Recent advances in nanomaterials integrated immunosensors for food toxin detection.

For the management and prevention of many chronic and acute diseases, the rapid quantification of toxicity in food and feed products have become a significant concern. Technology advancements in the area of biosensors, bioelectronics, miniaturization techniques, and microfluidics have shown a significant impact than conventional methods which have given a boost to improve the sensing performance towards food analyte detection. In this article, recent literature of Aflatoxin B1 (AFB1), worldwide permissible limits, major outbreaks and severe impact on healthy life have been discussed. An improvement achieved in detection range, limit of detection, shelf-life of the biosensor by integrated dimensional nanomaterials such as zero-dimension, one-dimension and two-dimension for AFB1 detection using electrical and optical transduction mechanism has been summarized. A critical overview of the latest trends using paper-based and micro-spotted array integrated with the anisotropic shape of nanomaterials, portable microfluidic devices have also been described together with future perspectives for further advancements.

Ultrasensitive Immunosensor Based on Langmuir-Blodgett Deposited Ordered Graphene Assemblies for Dengue Detection.

In this manuscript partially reduced graphene oxide (RGO) nanosheet-based electrodes have been utilized for quantification of the NS1 protein and subsequently for dengue detection. NS1 is the biomarker found circulating in the body of dengue-infected persons on or after first day of the appearance of disease symptoms. Graphene oxide (GO) has been synthesized using a modified Hummer's method, and its ordered nanostructured films have been electrophoretically deposited on indium tin oxide (ITO)-coated glass substrates using Langmuir-Blodgett (LB) deposition. Deposited LB films of GO have been reduced with hydrazine vapors to obtain RGO-coated ITO electrodes. NS1 antibodies have been grafted onto the ordered thin films using covalent linking, and the bioelectrodes have been utilized for the specific detection of NS1 antigen. The electrochemical performance of the fabricated bioelectrodes for NS1 antigen detection has been explored in standard and spiked sera samples. The limit of detection for the standard samples and spiked serum samples is found to be 0.069 ng mL-1 and 0.081 ng mL-1, respectively, with a sensitivity of 8.41 and 36.75 Ω per ng mL, respectively, in the detection range of 101 to 107 ng mL-1.

Gold nanobipyramids integrated ultrasensitive optical and electrochemical biosensor for Aflatoxin B1 detection.

This work reports the development of an electrical and optical biosensing for label-free detection of Aflatoxin B1 (AFB1) using gold (Au) nanobipyramids (NBPs). AuNBPs were synthesized through a two-step seed-mediated growth process followed by an exchange of capping agent from surfactant to lipoic acid. Pure and monodispersed AuNBPs of 70 nm base length were obtained and deposited on indium tin oxide (ITO)-coated glass substrate modified with self-assembled (3-Aminopropyl) triethoxysilane (APTES) film. The characterization of the obtained surfaces using spectroscopy, microscopy and diffractometry confirms the formation of AuNBPs, the conjugation to ITO electrode substrate and the immobilization of anti-AFB1 antibodies. AuNBPs modified ITO substrates were used for both electrochemical and Surface Plasmon Resonance biosensing studies. Localized Surface Plasmon Resonance (LSPR) local field enhancement was demonstrated. SPR based AFB1 detection was found to be linear in the 0.1-500 nM range with a limit of detection of 0.4 nM, whereas, impedimetric AFB1 detection was shown to be linear in the 0.1-25 nM range with a limit of detection of 0.1 nM. The practical utility of the impedimetric sensor was tested in spiked maize samples and 95-100% recovery percentage was found together with low relative standard deviation, proof of the robustness of this AFB1 sensor.

Integrated graphene quantum dot decorated functionalized nanosheet biosensor for mycotoxin detection.

Decoration of graphene quantum dots (GQDs) on molybdenum disulfide (MoS2) nanosheets serves as an active electrode material which enhances the electrochemical performance of the analyte detection system. Herein, ionic surfactant cetyltrimethylammonium bromide (CTAB)-exfoliated MoS2 nanosheets decorated with GQD material are used to construct an electrochemical biosensor for aflatoxin B1 (AFB1) detection. An antibody of AFB1 (aAFB1) was immobilized on the electrophoretically deposited MoS2@GQDs film on the indium tin oxide (ITO)-coated glass surface using a crosslinker for the fabrication of the biosensor. The immunosensing study investigated by the electrochemical method revealed a signal response in the range of 0.1 to 3.0 ng/mL AFB1 concentration with a detection limit of 0.09 ng/mL. Also, electrochemical parameters such as diffusion coefficient and heterogeneous electron transfer (HET) were calculated and found to be 1.67 × 10-5 cm2/s and 2 × 10-5 cm/s, respectively. The effective conjugation of MoS2@GQDs that provides abundant exposed edge sites, large surface area, improved electrical conductivity, and electrocatalytic activity has led to an excellent biosensing performance with enhanced electrochemical parameters. Validation of the fabricated immunosensor was performed in a spiked maize sample, and a good percentage of recoveries within an acceptable range were obtained (80.2 to 98.3%).Graphical abstract.

A label-free ultrasensitive microfluidic surface Plasmon resonance biosensor for Aflatoxin B1 detection using nanoparticles integrated gold chip.

The Surface Plasmon resonance (SPR) based label-free detection of small targeted molecules is a great challenge and require substantial signal amplification for the accurate and precise quantification. The incorporation of noble metal nanoparticles (NPs) like gold (Au) NPs for the fabrication of SPR biosensor has shown remarkable impact both for anchoring the signal amplification and generate plasmonic resonant coupling between NPs and chip surface. In this work, we present comparative studies related to the fabrication of self-assembled monolayer (SAM) and the influence of AuNPs on Au chip for Aflatoxin B1 (AFB1) detection using SPRi apparatus. The SAM Au chip was sequentially modified by EDC-NHS crosslinkers, grafting of protein-A and finally interaction with anti-AFB1 antibodies. Similar multilayer chip surface was prepared using functionalized lipoic acid AuNPs deposited on SAM Au chips followed by in situ activation of functional groups using EDC-NHS crosslinkers, grafting of protein-A and immobilization of anti-AFB1 antibodies. This multilayer functionalized AuNPs modified Au chip was successfully utilized for AFB1 detection ranging from 0.01 to 50 nM with a limit of detection of 0.003 nM. When compared to bare self-assembled Au chip which was shown to exhibit a limit of detection of 0.19 nM and a linear detection ranging from 1 to 50 nM, the AuNPs modified Au chip was proven to clearly be a better analytical tool. Finally, validation of the proposed biosensor was evaluated by spiked wheat samples and average recoveries (93 and 90.1%) were found to be acceptable.

Electrochemical Aflatoxin B1 immunosensor based on the use of graphene quantum dots and gold nanoparticles.

Electrochemical immunosensor for aflatoxin B1 (AFB1) is described that uses a composite prepared from graphene quantum dots (GQDs) and gold nanoparticles (Au NPs). The GQD-AuNP conjugate was obtained by using 2-aminothiophenol (ATP) as a linker where the carboxy groups of GQD bind to the amino groups of crosslinker via conjugation of thiol binding to the AuNP. To evaluate the conjugation of the GQD-AuNP composite, Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy (TEM) was applied. The composite was placed on an indium tin oxide (ITO) electrode and then modified with an antibody against AFB1. By using hexacyanoferrate as the electrochemical probe, the sensor works in the 0.1 to 3.0 ng mL-1 AFB1 concentration range, is highly specific, has good reproducibility and acceptable stability. The biosensor was applied to the analysis of (spiked) maize samples. Conceivably, the method can be utilized to sense other mycotoxins by using their respective antibodies. Graphical abstract Schematic presentation of electrochemical immunosensor for Aflatoxin B1 (AFB1) detection developed by antibodies of AFB1 (anti-AFB1) immobilization on graphene quantum dots (GQDs)-gold nanoparticles (AuNPs) composite deposited by electrophoretic deposition technique on an Indium tin oxide (ITO) surface.

Development of electrochemical biosensor based on CNT-Fe3O4 nanocomposite to determine formaldehyde adulteration in orange juice.

An electrochemical biosensor was developed to determine formaldehyde (HCHO) adulteration commonly found in food. The current responses of various electrodes based on multiwalled carbon nanotubes (CNTs) and synthesized nanocomposite (CNT-Fe3O4) were measured using cyclic voltammetry. The nanocomposite based biosensor shows comparatively high sensitivity (527 µA mg/L-1 cm-2), low detection limit (0.05 mg/L) in linear detection range 0.05-0.5 mg/L for formaldehyde detection using formaldehyde dehydrogenase (FDH) enzyme. In real sample analysis, the low obtained RSD values (less than 1.79) and good recovery rates (more than 90%) signify an efficient and precise sensor for the selective quantification of formaldehyde in orange juice. The developed biosensor has future implications for determining formaldehyde adulteration in citrus fruit juices and other liquid foods in agri-food chain to further resolve global food safety concerns, control unethical business practices of adulteration and reduce the widespread food borne illness outbreaks.

Highly efficient Polyaniline-MoS2 hybrid nanostructures based biosensor for cancer biomarker detection.

In this work, polyaniline nanospindles have been synthesized using iron oxide as sacrificial template. These nanospindles were utilized for the fabrication of PANI-MoS2 nanoflower architectures via hydrothermal route. The electrostatic interaction between PANI and MoS2 improves the conductivity and provides more direct paths for charge transportation. SEM, TEM, XRD, Raman Spectroscopy techniques were employed to explore the crystal structure, and morphological properties of the PANI-MoS2 nanocomposite. Furthermore, an electrochemical biosensing platform based on PANI-MoS2 nanocomposite was fabricated for the specific detection of chronic myelogenous leukemia (CML) by using electrochemical impedance spectroscopy technique. The binding interactions between the pDNA/PANI-MoS2/ITO bioelectrode and target DNA sequence were also studied. The biosensor exhibits high sensitivity and wide detection range (10-6  M to 10-17  M) of target DNA with low detection limit (3 × 10-18  M). Additionally, the specificity studies of the genosensor with various target DNA sequences (complementary, noncomplementary and one base mismatch) and real samples analysis of CML shows its potential for clinical diagnostics.

Graphene quantum dots-based nano-biointerface platform for food toxin detection.

Due to the similar electrochemical properties to graphene oxide (GO), graphene quantum dots (GQDs) are considered as a highly potential candidate for designing an electrochemical biosensor. In this report, GQDs were synthesized having an average diameter of 7 nm and utilized for the fabrication of an electrochemical immunosensor for the detection of food toxin, aflatoxin B1 (AFB1). An electrophoretic deposition technique was utilized to deposit the chemically synthesized GQDs onto indium tin oxide (ITO)-coated glass substrate. Further, the monoclonal antibodies of AFB1 were covalently immobilized onto deposited electrode GQDs/ITO using EDC-NHS as a crosslinker. The structural and morphological studies of GQDs and conjugated anti-AFB1 with GQDs have been investigated using UV-visible spectroscopy, photoluminescence spectroscopy, Raman spectroscopy, transmission electron microscopy, scanning electron microscopy techniques, etc. The electrochemical impedance spectroscopy and cyclic voltammetry measurements were carried out for electrical characterization and biosensing studies. This simple monodisperse GQDs-based platform yields heterogeneous electron transfer (97.63 × 10-5 cm s-1), the time constant (0.005 s) resulting in improved biosensing performance. The electrochemical immunosensor shows high sensitivity 213.88 Ω (ng mL-1)-1 cm-2. The limit of detection for standard samples and contaminated maize samples was found to be 0.03 ng mL-1 and 0.05 ng g-1, respectively, which is lower than the maximum acceptable limit according to the European Union. This result indicates its potential application for aflatoxin B1 detection in food contents. Graphical abstract ᅟ.

Electrochemical genosensor based on template assisted synthesized polyaniline nanotubes for chronic myelogenous leukemia detection.

This work reports a facile approach to synthesize polyaniline nanotubes (PANI-NT) by using manganese oxide as sacrificial templates. This template assisted polyaniline nanotubes (t-PANI-NT) were utilized as electrode material after deposition onto the indium tin oxide (ITO) coated glass substrates by using the electrophoretic technique. The structural, morphological and electrochemical characterizations of the t-PANI-NT show relatively better results compared to chemically synthesized PANI-NT (c-PANI-NT). Moreover, the t-PANI-NT/ITO electrode exhibits improved electron transfer coefficient (α = 0.63) and charge transfer rate constant (ks = 0.05912 s-1) in comparison to c-PANI-NT/ITO electrode (α = 0.56 and ks = 0.06548 s-1). The obtained t-PANI-NT/ITO electrodes have been further immobilized with biotinylated DNA sequence, specific to chronic myelogenous leukemia (CML) by using avidin-biotin as a cross-linking agent. Electrochemical impedance spectroscopy studies revealed that the genosensor displays linearity in wide range of target DNA concentration (10-6 to 10-16 M) with an outstanding differentiation ability and low detection limit of 10-16 M. The experimental results of this highly sensitive and specific genosensor with clinical samples of CML positive patients and control negative patients indicate its potential for clinical diagnostics.

Langmuir-Blodgett Nanoassemblies of the MoS2-Au Composite at the Air-Water Interface for Dengue Detection.

We report Langmuir-Blodgett (LB) films of molybdenum disulphide (MoS2) and gold nanoparticles (AuNP) composite being utilized as a biosensing platform for dengue detection. The LB films of the MoS2-AuNP composite have been transferred from the air-water interface to the indium tin oxide-coated glass substrate under optimized conditions. Further, antibodies specific to dengue NS1 antigen were immobilized onto these LB films. The fabricated immunosensor has been explored for NS1 antigen detection in standard samples as well as in spiked sera samples using electrochemical impedance spectroscopy. The NS1 antigen is present in the blood of infected persons from day one of the onset of clinical symptoms in primary dengue infection. The limit of detection for the standard and the spiked samples is found to be 1.67 and 1.19 ng mL-1, respectively, which is suitable for clinical applications, as NS1 antigen levels in patient's sera range from 0.04 to 2 µg mL-1 in primary infection and from 0.01 to 2 µg mL-1 in secondary infection.

Controlled deposition of functionalized silica coated zinc oxide nano-assemblies at the air/water interface for blood cancer detection.

We report results of the studies relating to controlled deposition of the amino-functionalized silica-coated zinc oxide (Am-Si@ZnO) nano-assemblies onto an indium tin oxide (ITO) coated glass substrate using Langmuir-Blodgett (LB) technique. The monolayers have been deposited by transferring the spread solution of Am-Si@ZnO stearic acid prepared in chloroform at the air-water interface, at optimized pressure (16 mN/m), concentration (10 mg/ml) and temperature (23 °C). The high-resolution transmission electron microscopic studies of the Am-Si@ZnO nanocomposite reveal that the nanoparticles have a microscopic structure comprising of hexagonal assemblies of ZnO with typical dimensions of 30 nm. The surface morphology of the LB multilayer observed by scanning electron microscopy shows uniform surface of the Am-Si@ZnO film in the nanometer range (<80 nm). These electrodes have been utilized for chronic myelogenous leukemia (CML) detection by covalently immobilizing the amino-terminated oligonucleotide probe sequence via glutaraldehyde as a crosslinker. The response studies of these fabricated electrodes carried out using electrochemical impedance spectroscopy show that this Am-Si@ZnO LB film based nucleic acid sensor exhibits a linear response to complementary DNA (10(-6)-10(-16) M) with a detection limit of 1 × 10(-16) M. This fabricated platform is validated with clinical samples of CML positive patients and the results demonstrate its immense potential for clinical diagnosis.

Thiol modified chitosan self-assembled monolayer platform for nucleic acid biosensor.

A self-assembled monolayer (SAM) of thiol modified chitosan (SH-CHIT), with thioglycolic acid (TGA) as a modifier to bestow thiol groups, has been prepared onto gold (Au)-coated glass plates for fabrication of the nucleic acid biosensor. The chemical modification of CHIT via TGA has been evidenced by Fourier transform infrared spectroscopy (FT-IR) studies, and the biocompatibility studies reveal that CHIT retains its biocompatible nature after chemical modification. The electrochemical studies conducted onto SH-CHIT/Au electrode reveal that thiol modification in CHIT amino end enhances the electrochemical behavior indicating that it may be attributed to delocalization of electrons in CHIT skeleton that participates in the resonance process. The carboxyl group modified end of DNA probe has been immobilized onto SH-CHIT/Au electrode using N-ethyl-N'-(3-dimethylaminopropyl)carbodimide (EDC) and N-hydroxysuccinimide (NHS) chemistry for detection of complementary, one-base mismatch and non-complementary sequence using electrochemical and optical studies for Mycobacterium tuberculosis detection. It has been found that DNA-SH-CHIT/Au bioelectrode can specifically detect 0.01 µM of target DNA concentration with sensitivity of 1.69 × 10(-6) A µM(-1).

Graphene oxide-based biosensor for food toxin detection.

We report results of the studies relating to the fabrication of a highly sensitive label free biosensor based on graphene oxide (GO) platform for the detection of aflatoxin B1 (AFB1) which is most toxic and predominant food toxin, using electrochemical impedance spectroscopy. The structural and optical characterization of GO/Au and anti-AFB1/GO/Au has been done by electron microscopy, Raman, X-ray diffraction (XRD), UV-vis and electrochemical impedance spectroscopy (EIS). The impedimetric sensing response of immunoelectrode as a function of AFB1 concentration reveals wider linear detection range (0.5-5 ng/ml), high sensitivity (639 Ω ng(-1) ml), improved detection limit (0.23 ng ml(-1)) and good stability (5 weeks) for the label-free detection. Association constant (k a) for antigen-antibody interaction obtained as 0.46 ng ml(-1) indicates high affinity.

Nanostructuring of hierarchical 3D cystine flowers for high-performance electrochemical immunosensor.

Here, we report a simple and reproducible method for large scale fabrication of novel flower and palm-leaf like 3D cystine microstructures (CMs) with high uniformity having a size of 50 µm and 10 µm respectively, through a facile aqueous solution route as a function of pH and concentration. In a proof-of-concept study, the 3D CMs have been further explored to fabricate a label-free high-performance electrochemical immunosensor by immobilizing monoclonal antibodies. Electrochemical methods were employed to study the stepwise modification of the system and the electronic transduction for the detection. The fabricated immunosensor design demonstrates high performance with enhanced sensitivity (4.70 cfu ml(-1)) and linear sensing range from 10 to 3 x 10(9) cfu ml(-1) a long shelf-life (35 days) and high selectivity over other bacterial pathogens. The enhanced performance originates from a novel nanostructuring in which the CMs provide higher surface coverage for the immobilization of antibodies providing excellent electronic/ionic conductivity which result in the enhanced sensitivity.

Coupling electrochemical response of a DNA biosensor with PCR for Neisseria gonorrhoeae detection.

Early diagnosis of gonococcal infections is important with regard to a patient's health and stage of infection. In this context, we report the development of an opa-gene-based electrochemical DNA biosensor for detection of Neisseria gonorrhoeae by monitoring redox peak of methylene blue indicator. The fabricated biosensor has been shown to be highly sensitive and specific when evaluated with complementary, non-complementary, and 1-base mismatch DNA sequences and polymerase chain reaction (PCR) amplified products (amplicons) of standard strain of N. gonorrhoeae (ATCC49226). The biosensor has been further evaluated using amplicons of known positive and negative clinical samples, and cut-off for positives has been determined using receiver operating characteristic curve. The sensitivity (SN), specificity (SP), positive predictive value, and negative predictive value of the biosensor have been found to be 96.2%, 88.2%, 92.6%, and 93.8%, respectively. We conclude that the combination of PCR amplification with electrochemical detection shows distinct advantage of high SN and increased SP for gonococcal detection.

Quantum dots self assembly based interface for blood cancer detection.

Results of the studies related to fabrication of sensitive electrochemical biosensor using an interface based on quantum dots (QDs) self-assembly is reported. The QDs assembly is sought to provide improved fundamental characteristics to the electrode interface in terms of electroactive surface area, diffusion coefficient, and electron transfer kinetics. This QDs modified electrode has been utilized to serve as a transducer surface for covalent immobilization of chronic myelogenous leukemia (CML) specific probe oligonucleotide, designed from the BCR-ABL fusion gene. The electrochemical characteristics of this biosensor toward various designed synthetic oligonucleotides reveal a significant enhancement in its mismatch discrimination capability compared to the biosensing assay without QDs under similar experimental conditions. The sensing characteristics of this biosensor offer a potential for detection of target oligonucleotide at a concentration as low as 1.0 pM. Furthermore, the PCR-amplified CML-positive patient samples with various BCR-ABL transcript ratios can be electrochemically distinguished from healthy samples, indicating promising application of the QDs based biosensor for clinical investigations.

Electrophoretically deposited reduced graphene oxide platform for food toxin detection.

Reduced graphene oxide (RGO) due to its excellent electrochemical properties and large surface area, has recently aroused much interest for electrochemical biosensing application. Here, the chemically active RGO has been synthesized and deposited onto an indium tin oxide (ITO) coated glass substrate by the electrophoretic deposition technique. This novel platform has been utilized for covalent attachment of the monoclonal antibodies of aflatoxin B1 (anti-AFB1) for food toxin (AFB1) detection. The electron microscopy, X-ray diffraction, and UV-visible studies reveal successful synthesis of reduced graphene oxide while the XPS and FTIR studies suggest its carboxylic functionalized nature. The electrochemical sensing results of the anti-AFB1/RGO/ITO based immunoelectrode obtained as a function of aflatoxin concentration show high sensitivity (68 µA ng(-1) mL cm(-2)) and improved detection limit (0.12 ng mL(-1)). The association constant (ka) for antigen-antibody interaction obtained as 5 × 10(-4) ng mL(-1) indicates high affinity of antibodies toward the antigen (AFB1).