Abstract

In the present article, we developed a highly sensitive label-free electrochemical immunosensor based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO)/gold nanoparticles modified glassy carbon electrode for the determination of receptor tyrosine kinase-like orphan receptor (ROR)-1. In this electrochemical immunoassay platform, NiFe-LDH/rGO was used due to great electron mobility, high specific surface area and flexible structures, while Au nanoparticles were prepared and coated on the modified electrodes to improve the detection sensitivity and ROR1 antibody immobilizing (ROR1Ab). The modification procedure was approved by using cyclic voltammetry and differential pulse voltammetry based on the response of peak current to the step by step modifications. Under optimum conditions, the experimental results showed that the immunosensor revealed a sensitive response to ROR1 in the range of 0.01–1 pg mL−1, and with a lower limit of quantification of 10 attogram/mL (10 ag mL−1). Furthermore, the designed immunosensor was applied for the analysis of ROR1 in several serum samples of chronic lymphocytic leukemia suffering patients with acceptable results, and it also exhibited good selectivity, reproducibility and stability.

Highlights

  • The receptor tyrosine kinase-like orphan receptor 1 (ROR1), an oncofetal antigen, is a Wnt5a receptor and is widely expressed during embryogenesis, whereas it is absent in most adult ­tissues[1,2]

  • A new label-free electrochemical platform based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO) nanocomposites and AuNPs was designed and constructed for the detection of ROR1 biomarker

  • A stable NiFe-LDH/rGO nanocomposite with appropriate and acceptable electrocatalytic activity was first electrodeposited on the glassy carbon electrode (GCE)

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Summary

Introduction

The receptor tyrosine kinase-like orphan receptor 1 (ROR1), an oncofetal antigen, is a Wnt5a receptor and is widely expressed during embryogenesis, whereas it is absent in most adult ­tissues[1,2]. The poor conductivity of LDH often limits electron transfer; so, their electrochemical activity can be enhanced excellently when they are combined with nano-carbon ­materials[26]. A new label-free electrochemical platform based on NiFe-LDH/rGO nanocomposites and AuNPs was designed and constructed for the detection of ROR1 biomarker. Label free electrochemical immunosensor with high sensitivity can detect very low amount of ROR1 and help in the point of care diagnosis of most cancers. It will be more cost-effective and simpler than conventional ROR1 detection methods. In other word the incubation time of secondary antibody with the target protein is neglected

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