Multiplexed biosensing methods for simultaneously detecting multiple biomolecules are important for investigating biological mechanisms associated with physiological processes, developing applications in life sciences, and conducting medical tests. The development of biosensors, especially those advanced biosensors with multiplexing potentials, strongly depends on advancements in nanotechnologies, including the nano-coating of thin films, micro–nano 3D structures, and nanotags for signal generation. Surface functionalization is a critical process for biosensing applications, one which enables the immobilization of biological probes or other structures that assist in the capturing of biomolecules. During this functionalizing process, nanomaterials can either be the objects of surface modification or the materials used to modify other base surfaces. These surface-functionalizing strategies, involving the coordination of sensor structures and materials, as well as the associated modifying methods, are largely determinative in the performance of biosensing applications. This review introduces the current studies on biosensors with multiplexing potentials and focuses specifically on the roles of nanomaterials in the design and functionalization of these biosensors. A detailed description of the paradigms used for method selection has been set forth to assist understanding and accelerate the application of novel nanotechnologies in the development of biosensors.

Background: A central issue hindering the development of effective anti-fibrosis drugs for heart failure is the unclear interrelationship between fibrosis and the immune cells. This study aims at providing precise subtyping of heart failure based on immune cell fractions, elaborating their differences in fibrotic mechanisms, and proposing a biomarker panel for evaluating intrinsic features of patients’ physiological statuses through subtype classification, thereby promoting the precision medicine for cardiac fibrosis.Methods: We inferred immune cell type abundance of the ventricular samples by a computational method (CIBERSORTx) based on ventricular tissue samples from 103 patients with heart failure, and applied K-means clustering to divide patients into two subtypes based on their immune cell type abundance. We also designed a novel analytic strategy: Large-Scale Functional Score and Association Analysis (LAFSAA), to study fibrotic mechanisms in the two subtypes.Results: Two subtypes of immune cell fractions: pro-inflammatory and pro-remodeling subtypes, were identified. LAFSAA identified 11 subtype-specific pro-fibrotic functional gene sets as the basis for personalised targeted treatments. Based on feature selection, a 30-gene biomarker panel (ImmunCard30) established for diagnosing patient subtypes achieved high classification performance, with the area under the receiver operator characteristic curve corresponding to 0.954 and 0.803 for the discovery and validation sets, respectively.Conclusion: Patients with the two subtypes of cardiac immune cell fractions were likely having different fibrotic mechanisms. Patients’ subtypes can be predicted based on the ImmunCard30 biomarker panel. We envision that our unique stratification strategy revealed in this study will unravel advance diagnostic techniques for personalised anti-fibrotic therapy.