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This work demonstrates the ability of the Ion-Sensitive Field-Effect Transistor (ISFET)-based immunosensor to detect antibodies against the human leukocyte antigen (HLA) and the major histocompatibility complex class-I-related chain A (MICA). The sensing membrane of the ISFET devices was modified and functionalized using an APTES-GA strategy. Surface properties, including wettability, surface thickness, and surface topology, were assessed in each module of the modification process. The optimal concentrations of HLA and MICA proteins for the immobilization were 10 and 50 μg/mL. The dose-response curve showed a detection range of 1.98-40 µg/mL for anti-HLA and 5.17-40 µg/mL for anti-MICA. The analytical precision (%CV) was found to be 10.69% and 8.92% for anti-HLA and -MICA, respectively. Moreover, the electrical signal obtained from the irrelevant antibody was considerably different from that of the specific antibodies, indicating the specific binding of the relevant antibodies without noise interference. The sensitivity and specificity in the experimental setting were established for both antibodies (anti-HLA: sensitivity = 80.00%, specificity = 86.36%; anti-MICA: sensitivity = 86.67%, specificity = 88.89%). Our data reveal the potential of applying the ISFET-based immunosensor to the detection of relevant anti-HLA and -MICA antibodies, especially in the field of kidney transplantation.

In addition to their use as an additive to improve physical properties of solvent polymeric membranes, plasticizers have a considerable impact on the specificity and sensitivity of membrane-modified electrochemical sensors. In this work, we aim at the hybridization of two different plasticizers using the electropolymerization technique in the development of a cadmium(II)-selective electrochemical sensor based on screen-printed gold electrode along with cyclic voltammetric measurement. At this point, we first screen for the primary plasticizer yielding the highest signal using cyclic voltammetry followed by pairing it with the secondary plasticizers giving rise to the most sensitive current response. The results show that the hybridization of DOS and TOTM with 3:1 weight ratio (~137.7-μm-thick membrane) renders a signal that is >26% higher than that from the sensor plasticized by DOS per se in water. The solution of 0.1 mM hydrochloric acid (pH 4) is the optimal supporting electrolyte. In addition, hybrid plasticizers have adequate redox capacity to induce cadmium(II) transfer from bulk solution to the membrane/water interfaces. Conversion of voltammetric signals to semi-integral currents results in linearity with cadmium(II) concentration, indicating the irreversible cadmium(II) transfer to the membrane. The DOS:TOTM hybrid sensor also exhibits high sensitivity, with a limit of detection (LOD) and limit of quantitation (LOQ) of 95 ppb and 288 ppb, respectively, as well as greater specificity towards cadmium(II) than that obtained from the single plasticizer sensor. Furthermore, recovery rates of spiked cadmium(II) in water samples were higher than 97% using the hybrid plasticizer sensor. Unprecedentedly, our work reports that the hybridization of plasticizers serves as ion-to-electron transducer that can improve the sensor performance in cadmium(II) detection.

The 3-aminopropyltriethoxysilane (APTES) is a common method for biomolecule immobilization on silicon and silicon derivatives such as silicon nitride (Si 3 N 4 ). However, there are many parameters which impact the efficiency of APTES modification such as APTES concentration and reaction time. Thus, various APTES concentrations (0.1%, 0.5%, 1%, 2%, 5%, and 10%) under different reaction times (15, 30, 60 and 120 min) were compared to achieve the optimal APTES modification condition which produced a thin and stable APTES layer on Si 3 N 4 surface. The modified surfaces were characterized by contact angle (CA) measurement, Fourier transform infrared (FTIR) spectroscopy and spectroscopic ellipsometry to determine the wetting property, chemical bonding composition and surface thickness, respectively. In addition, biotin was used as a model to determine the effectiveness of APTES modification condition by coupling with glutaraldehyde (GA). The Alexa Flour 488 conjugated streptavidin was performed to visualize the presence of biotin using fluorescence microscopy due to the specifically binding between biotin and streptavidin. The atomic force microscopy (AFM) was utilized to determine the surface topology which was an indicator to demonstrate the agglomeration of APTES molecule. Moreover, ion sensitive field effect transistor (ISFET) was employed as a biosensor model to demonstrate the effect between surface thickness and sensitivity of biosensor. The results show that the APTES thickness is directly correlated to the APTES concentration and reaction time. Since the importance parameter for ISFET measurement is the distance between biomolecule and sensing membrane of ISFET, the thicker APTES layer negatively impacts the sensitivity of ISFET based biosensor because of the ion shielding effect. Therefore, these results would be valuable information for development of Si 3 N 4 biosensor, especially ISFET based biosensor., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A silicon nitride Ion Sensitive Field Effect Transistor (ISFET) based immunosensor was developed as a low-cost and label-free electrical detection for the detection of antigen 85 complex B (Ag85B). The sensing membrane of the ISFET was modified with 3-aminopropyltriethoxysilane (APTES) followed by glutaraldehyde (GA), yielding an aldehyde-terminated surface. This group is available for immobilization of a monoclonal antibody against a recombinant Ag85B protein (anti-Ag85B antibody). The optimal concentration for anti-Ag85B antibody immobilization onto the modified ISFET was 100 μg ml -1 . This optimal condition provided the maximal binding capability and minimal non-specific background signal. The binding event between the recombinant Ag85B antigen and anti-Ag85B antibody on the ISFET surface is presented by monitoring the gate potential change at a constant drain current. The dose response for the recombinant Ag85B protein showed a linear response between 0.12 and 1 μg ml -1 without significant interference from other recombinant proteins. The analytical imprecision (CV%) and accuracy of this Ag85B protein biosensor were 9.73-10.99% and 95.29%, respectively. In addition, an irrelevant antibody and other recombinant proteins were employed as a negative control to demonstrate the non-specific interaction of the antigen and antibody. The success of this immunosensor system for Ag85B protein detection facilitates the construction of a promising device which can shorten the turnaround time for the diagnosis of tuberculosis compared to a standard culture method. Furthermore, this device could also be applied for real-time growth monitoring of Mycobacterium tuberculosis in a mycobacterial culture system.

Three different types of surface, silicon dioxide (SiO2), silicon nitride (Si3N4), and titanium oxynitride (TiON) were modified for lactate dehydrogenase (LDH) immobilization using (3-aminopropyl)triethoxysilane (APTES) to obtain an amino layer on each surface. The APTES modified surfaces can directly react with LDH via physical attachment. LDH can be chemically immobilized on those surfaces after incorporation with glutaraldehyde (GA) to obtain aldehyde layers of APTES-GA modified surfaces. The wetting properties, chemical bonding composition, and morphology of the modified surface were determined by contact angle (CA) measurement, Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM), respectively. In this experiment, the immobilized protein content and LDH activity on each modified surface was used as an indicator of surface modification achievement. The results revealed that both the APTES and APTES-GA treatments successfully link the LDH molecule to those surfaces while retaining its activity. All types of tested surfaces modified with APTES-GA gave better LDH immobilizing efficiency than APTES, especially the SiO2 surface. In addition, the SiO2 surface offered the highest LDH immobilization among tested surfaces, with both APTES and APTES-GA modification. However, TiON and Si3N4 surfaces could be used as alternative candidate materials in the preparation of ion-sensitive field-effect transistor (ISFET) based biosensors, including lactate sensors using immobilized LDH on the ISFET surface., (Copyright © 2014 Elsevier B.V. All rights reserved.)