The detection of alpha-fetoprotein (AFP) in plasma is important in the diagnosis of hepatocellular carcinoma (HCC) in humans. We developed a biosensor to detect AFP in HCC patient plasma and in a phosphate buffer saline (PBS) solution using a graphene field-effect transistor (G-FET). The G-FET was functionalized with 1-pyrenebutyric acid N-hydroxysuccinimide ester (PBASE) for immobilization of an anti-AFP antibody. AFP was detected by assessing the shift in the voltage of the Dirac point (ΔVDirac) after binding of AFP to the anti-AFP-immobilized G-FET channel surface. This anti-AFP-immobilized G-FET biosensor was able to detect AFP at a concentration of 0.1 ng mL−1 in PBS, and the detection sensitivity was 16.91 mV. In HCC patient plasma, the biosensor was able to detect AFP at a concentration of 12.9 ng mL−1, with a detection sensitivity of 5.68 mV. The sensitivity (ΔVDirac) depended on the concentration of AFP in either PBS or HCC patient plasma. These data suggest that G-FET biosensors could have practical applications in diagnostics.
A reference electrode is necessary for the working of ion-sensitive field-effect transistor (ISFET)-type sensors in electrolyte solutions. The Ag/AgCl electrode is normally used as a reference electrode. However, the Ag/AgCl reference electrode limits the advantages of the ISFET sensor. In this work, we fabricated a two-channel graphene solution gate field-effect transistor (G-SGFET) to detect pH without an Ag/AgCl reference electrode in the electrolyte solution. One channel is the sensing channel for detecting the pH and the other channel is the reference channel that serves as the reference electrode. The sensing channel was oxygenated, and the reference channel was fluorinated partially. Both the channels were directly exposed to the electrolyte solution without sensing membranes or passivation layers. The transfer characteristics of the two-channel G-SGFET showed ambipolar field-effect transistor (FET) behavior (p-channel and n-channel), which is a typical characteristic curve for the graphene ISFET, and the value of VDirac was shifted by 18.2 mV/pH in the positive direction over the range of pH values from 4 to 10. The leakage current of the reference channel was 16.48 nA. We detected the real-time pH value for the two-channel G-SGFET, which operated stably for 60 min in the buffer solution.
The monitoring of Na+ ions distributed in the body has been indirectly calculated by the detection of Na+ ions in urine. We fabricated a two–dimensional (2D) Na+ ion sensor using a graphene ion–sensitive field–effect transistor (G–ISFET) and used fluorinated graphene as a reference electrode (FG–RE). We integrated G–ISFET and FG on a printed circuit board (PCB) designed in the form of a secure digital (SD) card to fabricate a disposable Na+ ion sensor. The sensitivity of the PCB tip to Na+ ions was determined to be −55.4 mV/dec. The sensor exhibited good linearity despite the presence of interfering ions in the buffer solution. We expanded the evaluation of the PCB tip to real human patient urine samples. The PCB tip exhibited a sensitivity of −0.36 mV/mM and linearly detected Na+ ions in human patient urine without any dilution process. We expect that G–ISFET with FG–RE can be used to realize a disposable Na+ ion sensor by serving as an alternative to Ag/AgCl reference electrodes.
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