17β-estradiol (17β-E2) plays a critical role in regulating reproduction in human, there is therefore an urgent need to detect it sensitively and precisely in a cost-effective and easy method. In this paper, a label-free integrated microfluidic paper-based analytical device was developed for highly sensitive electrochemical detection of 17β-E2. The microfluidic channel of the paper-based sensor was fabricated with wax printing and the three electrodes, including working, counter and reference electrode were screen-printed. Multi-walled carbon nanotubes (MWCNTs)/ thionine (THI)/ gold nanoparticles (AuNPs) Nano composites were synthesized and coated on screen-printed working electrode (SPWE) for the immobilization of anti-E2. In this electro-chemical system of paper-based immunoassay, THI molecules serving as an electrochemical mediator while MWCNTs and AuNPs, due to their excellent electrical conductivities, could accelerate electron transfer for the signal amplification. Experimental results revealed that the immunoassay is able to detect 17β-E2 as low as 10 pg mL-1, with a linear range from 0.01 to 100 ng mL-1. This microfluidic paper-based immunosensor would provide a new platform for low cost, sensitive, specific, and point-of-care diagnosis of 17β-E2.

In this work, an electrochemical paper-based aptasensor was fabricated for label-free and ultrasensitive detection of epidermal growth factor receptor (EGFR) by employing anti-EGFR aptamers as the bio-recognition element. The device used the concept of paper-folding, or origami, to serve as a valve between sample introduction and detection, so reducing sampling volumes and improving operation convenience. Amino-functionalized graphene (NH₂-GO)/thionine (THI)/gold particle (AuNP) nanocomposites were used to modify the working electrode not only to generate the electrochemical signals, but also to provide an environment conducive to aptamer immobilization. Electrochemical characterization revealed that the formation of an insulating aptamer–antigen immunocomplex would hinder electron transfer from the sample medium to the working electrode, thus resulting in a lower signal. The experimental results showed that the proposed aptasensor exhibited a linear range from 0.05 to 200 ngmL⁻¹ (R² = 0.989) and a detection limit of 5 pgmL⁻¹ for EGFR. The analytical reliability of the proposed paper-based aptasensor was further investigated by analyzing serum samples, showing good agreement with the gold-standard enzyme-linked immunosorbent assay.