Electrochemical Biosensing Strategy for Ultrasensitive Detection of Aquaporin-4 Antibody as Neuromyelitis Optica Autoimmune Disease Biomarker Using Aquaporin-4 Extracellular Loop

Abstract

Neuromyelitis optica (NMO) is a severe and disabling neurodegenerative disorder of the central nervous system (CNS). Neuromyelitis optica-Immunoglobulin G (NMO-IgG) is a serum IgG autoantibody almost exclusively present in NMO patients, which helps to differentiate NMO from other CNS disorders. Developing standardized and user-friendly assays remains a significant challenge in making NMO-IgG testing widely available. Label-free methods are simpler and faster, without additional reagents and procedures. Here, we present a peptide-based label-free electrochemical biosensor for detecting aquaporin-4 antibodies (AQP4-Abs) using extracellular AQP4 to diagnose NMO disease via the DPV electrochemical method. We have developed a novel approach in which the E loop of extracellular AQP4 is bemployed to detect NMO. 3 phenylalanines (Phe) were annexed to the C terminal, and because phenylalanine has a benzene ring, it can have π-π interaction with the benzene ring of carbon nanotube (CNT). In the designated platform, instead of using functional groups with complex and multi-step processes for immobilizing on the electrode surface, we used Nickel-Metal−organic framework /CNT as a novel modifier for measuring AQP4 antibodies with a simple, cheap, and accessible synthesis method. The developed sensor can detect antibodies with detection limit and quantification of 6.2 and 10.0 pg ml−1, respectively (S/N = 3). Also, superb sensitivity of the biosensor was attained as 28.8 μA mL ng−1 cm−2, confirming that the sensor has great potential for clinical application as a diagnostic test.