Matrix tolerance, probe orientation, and antifouling interfaces in graphene biosensors

Graphene-based biosensors offer high analytical sensitivity, positioning graphene as a leading transducer material for next-generation environmental, food-safety, and biomedical diagnostic applications. As the field advances toward real-world implementation, maintaining performance in complex biological and environmental matrices remains a central challenge. Under these conditions, analytical reliability is governed not only by graphene's intrinsic properties, but also by the structural and functional organization of the bio-nano interface. This review examines recent interface-engineering strategies that address these challenges through three interconnected design parameters: matrix tolerance, probe orientation, and antifouling. We discuss how porous and nanostructured interfaces regulate mass transport and preserve signal integrity in minimally processed samples, how linker chemistry and surface architecture control the accessibility and presentation of biorecognition elements within the effective transduction zone, and how hydrated antifouling layers suppress nonspecific adsorption while stabilizing interfacial charge transfer. We further discuss multiparametric interface engineering as a unifying framework for improving selectivity, robustness, quantitative accuracy, and sensitivity. The review highlights critical design principles, benchmarking needs, and translational priorities for the development of graphene biosensors intended for clinical, environmental, and food-monitoring applications.