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Objective: This review aims to make a brief overview of blood biomarkers’ clinical decision limits, possibly aiding in outcome prediction in all-aged Tetralogy of Fallot patients. Potentially, these biomarkers could also assist in necessity and timing of pulmonary valve replacement.

The reliability of analysis is becoming increasingly important as point-of-care diagnostics are transitioning from single-analyte detection toward multiplexed multianalyte detection. Multianalyte detection benefits greatly from complementary metal-oxide semiconductor (CMOS) integrated sensing solutions, offering miniaturized multiplexed sensing arrays with integrated readout electronics and extremely large sensor counts. The development of CMOS back end of line integration compatible graphene field-effect transistor (GFET)-based biosensing has been rapid during the past few years, in terms of both the fabrication scale-up and functionalization toward biorecognition from real sample matrices. The next steps in industrialization relate to improving reliability and require increased statistics. Regarding functionalization toward truly quantitative sensors, on-chip bioassays with improved statistics require sensor arrays with reduced variability in functionalization. Such multiplexed bioassays, whether based on graphene or on other sensitive nanomaterials, are among the most promising technologies for label-free electrical biosensing. As an important step toward that, we report wafer-scale fabrication of CMOS-integrated GFET arrays with high yield and uniformity, designed especially for biosensing applications. We demonstrate the operation of the sensing platform array with 512 GFETs in simultaneous detection for the sodium chloride concentration series. This platform offers a truly statistical approach on GFET-based biosensing and further to quantitative and multianalyte sensing. The reported techniques can also be applied to other fields relying on functionalized GFETs, such as gas or chemical sensing or infrared imaging., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

We demonstrate a label-free biosensor concept based on specific receptor modules, which provide immobilization and selectivity to the desired analyte molecules, and on charge sensing with a graphene field effect transistor. The receptor modules are fusion proteins in which small hydrophobin proteins act as the anchor to immobilize the receptor moiety. The functionalization of the graphene sensor is a single-step process based on directed self-assembly of the receptor modules on a hydrophobic surface. The modules are produced separately in fungi or plants and purified before use. The modules form a dense and well-oriented monolayer on the graphene transistor channel and the receptor module monolayer can be removed, and a new module monolayer with a different selectivity can be assembled in situ. The receptor module monolayers survive drying, showing that the functionalized devices can be stored and have a reasonable shelf life. The sensor is tested with small charged peptides and large immunoglobulin molecules. The measured sensitivities are in the femtomolar range, and the response is relatively fast, of the order of one second.

We present an approach where biomolecular self-assembly is used in combination with lithography to produce patterns of metallic nanoparticles on a silicon substrate. This is achieved through a two-step method, resulting in attachment of nanoparticles on desired sites on the sample surfaces, which allowed a detailed characterization. First, a genetically modified hydrophobin protein, NCysHFBI, was attached by self-assembly on a hydrophobic surface or a surface patterned with hydrophobic and hydrophilic domains. The next step was to label the protein layers with 17.8 nm gold nanoparticles, to allow microscopic characterization of the films. Kinetics and extent of attachment of nanoparticles were characterized by UV-vis spectroscopy and transmission electron microscopy. It was shown that the attachment of citrate-stabilized gold nanoparticles was strongly dependent on the electrostatic properties of the capping ligand layer and the density of nanoparticles in the monolayer could be controlled via pH. The resulting nanoparticle assemblies followed the original pattern created by optical lithography in high accuracy. We demonstrate that combining bottom-up and top-down nanotechnological approaches in a good balance can provide very effective ways to produce nanoscale components providing a functional interface between electronics and the biological world.