Lab-on-paper aptasensor for label-free picomolar detection of a pancreatic hormone in serum

Abstract

Pancreatic hormones such as insulin play the crucial life-supporting role of glucose metabolism. Due to rapidly growing diabetic disorders and associated health complications globally, combined with the pathogenic viral infections severely affecting people with pre-existing conditions, new user-friendly, affordable molecular diagnostic approaches that offer simplicity, cost-effectiveness, and an ultra-sensitive detection in human biofluids are necessary to improve the global health. In this regard, a low-cost paper device that enables easy monitoring of fasting blood-relevant ultra-low picomolar insulin levels is innovative, giving broader applications to any other similar critical molecular targets. The essential contribution of this lab-on-paper is the combination of label-free electrochemical insulin aptasensing with a paper electrode device as a simpler, cheaper, and reliable molecular diagnostic approach for complex serum samples with supporting independent validation methods for confirming scientific rigor and fit-for-purpose of the presented approach. Our aptasensor features a carboxylated graphene-aptamer surface design offering a clinically relevant quantitative detection of picomolar insulin present in a 10-fold diluted neat serum based on interfacial capacitance changes in proportion to the serum insulin concentration (dynamic range 5–500 pM and limit of detection 1.5 pM). Application for undiluted serum samples is demonstrated. Furthermore, analysis of an actual diabetes patient serum sample and correlation of the capacitance sensor results with peroxidase antibody label-based insulin assay approaches are presented (amperometric detection and a commercial enzyme-linked immunosorbent assay are used as validation methods).