Facilitated detection of RNA N6-Methyladenosine via EDC/NHS-protein a functionalized laser-induced graphene electrodes

N6-methyladenosine (m6A) is the most prevalent epigenetic RNA modification in eukaryotes, playing critical roles in RNA metabolism and disease progression. Current detection methods for m6A, such as mass spectrometry and antibody-based assays, face limitations in sensitivity, cost, or complexity. Electrochemical biosensors offer a promising alternative but struggle with nonspecific binding and low signal-to-noise ratios. This work addresses these challenges by developing a highly sensitive and selective electrochemical sensor for m6A quantification in complex biological samples. We designed a laser-induced graphene electrode (AuNPs/LIG) functionalized with gold nanoparticles and 11-mercaptoundecanoic acid (MUA) to immobilize anti-m6A antibodies via Protein A (PrA)-directed coupling. The sensor exploited steric hindrance and electrostatic repulsion from m6A-bound RNA to suppress electron transfer of the Fe[(CN) 6 ] 3− redox probe, yielding a current response proportional to m6A concentration. The assay achieved a linear range of 0.001–10 nM with a detection limit of 0.284 pM, surpassing our previous work by a large margin in terms of sensitivity. Validation in HeLa cell RNA and undiluted human serum confirmed high accuracy and minimal interference from nonspecific RNA. This method of m6A detection offers unparalleled advantages in sensitivity, real-time quantification, and cost-effective miniaturization, while future advancements in material science and multiplexed detection integration could revolutionize its clinical translation for point-of-care diagnostics in m6A-related diseases, such as cancer and metabolic disorders.