Binary MoS2 nanostructures as nanocarriers for amplification in multiplexed electrochemical immunosensing: simultaneous determination of B cell activation factor and proliferation-induced signal immunity-related cytokines

Beatriz Arévalo,Alejandro Valverde,Paloma Yáñez-Sedeño,Susana Campuzano,Guillermo Solís-Fernández,Rodrigo Barderas,Verónica Serafín,José M Pingarrón,Marina Blázquez-García,Ana Montero-Calle

doi:10.1007/s00604-022-05250-4

Abstract

A dual immunosensor is reported for the simultaneous determination of two important immunity-related cytokines: BAFF (B cell activation factor) and APRIL (a proliferation-induced signal). Sandwich-type immunoassays with specific antibodies (cAbs) and a strategy for signal amplification based on labelling the detection antibodies (dAbs) with binary MoS2/MWCNTs nanostructures and using horseradish peroxidase (HRP) were implemented. Amperometric detection was carried out at screen-printed dual carbon electrodes (SPdCEs) through the hydroquinone HQ/H2O2 system. The developed dual immunosensor provided limit of detection (LOD) of 0.08 and 0.06 ng mL−1 for BAFF and APRIL, respectively, and proved to be useful for the determination of both cytokines in cancer cell lysates and serum samples from patients diagnosed with autoimmune diseases and cancer. The obtained results agreed with those found using ELISA methodologies.Graphical abstract

Highlights

BAFF (B cell activating factor) and APRIL are molecules similar to tumor necrosis factor mainly expressed on B lymphocytes that show structural similarities and interact with three receptors of the TNF family and redundantly: BAFF-R joins BAFF, BCMA joins APRIL and BAFF with weaker affinity, and TACI joins BAFF and APRIL well [1]
500 μL of 0.1 mg mL−1 MoS2/ Multi-walled carbon nanotubes (MWCNTs) was centrifuged at 14,000 rpm during 10 min and the remaining product was incubated in the darkness under continuous stirring for 6 h at room temperature with 200 μL of a 400 mM EDC and 100 mM NHSS mixture solution prepared in 100 mM phosphate buffer saline solution (PBS) pH 7.4
MoS2 composites prepared with carbon nanotubes mostly maintain the bare material structure providing rough surface morphology in the form of nanoaggregates, while in the presence of graphene oxide (GO), the synthesized M oS2 appears as welldispersed sheets on rGO substrate

Summary

Introduction

BAFF (B cell activating factor) and APRIL (a proliferationinducing ligand) are molecules similar to tumor necrosis factor mainly expressed on B lymphocytes that show structural similarities and interact with three receptors of the TNF family and redundantly: BAFF-R joins BAFF, BCMA joins APRIL and BAFF with weaker affinity, and TACI joins BAFF and APRIL well [1] Both are type II membrane proteins, and the former is released after being cleaved at the furin protease site as a biologically active and soluble 17 kDa cytokine [2]. Sandwich-type immunoassays were implemented using M oS2/ MWCNTs(-HRP)-dAbs as carrier labels for signal amplification In these nanocarriers the acid-treated MWCNTs provided the required functional groups to immobilize a large number of HRP and dAb molecules, and MoS2 contributed to improve sensitivity due to its pseudo-peroxidase activity. ELISA absorbance readings were made in a SunriseTM Tecan microplate reader with the Magellan V 7.1 software

Procedures

Results and discussion

Conclusions