RESUMO
A ratiometric electrochemical immunosensor is proposed for simultaneous detection of cellular-myelocytomatosis oncoprotein (C-myc) and B-cell lymphoma 2 (Bcl-2) via the potential-resolved strategy. It relied on multi-role co-loaded alloy composites (CLACs) and poly(3,4-ethylenedioxythiophene) (PEDOT)-graphene oxide (GO)-multiwalled carbon nanotubes (MWCNTs) (PGM) modified electrodes. CLACs with good catalytic and enzyme-like properties were synthesized in one step by loading tetramethylbenzidine (TMB) or methylene blue (MB) into Pt-Pd alloy and used as label materials. After immunological reactions, CLACs showed distinguishable dual differential pulse voltammetry signals at - 0.26 V and 0.38 V, corresponding to C-myc and Bcl-2, and the PGM had an electrochemical signal at 1.2 V, which could be used as a reference signal to construct a ratiometric sensor. CLACs had a satisfactory synergistic effect with the PGM, and eventually achieved quadruple signal amplification. Thus, benefiting from multiple magnification and ratiometric self-calibration functions, sensitive detections of C-myc and Bcl-2 were achieved, with detection limits as low as 0.5 and 2.5 pg mL-1, respectively. Additionally, when the designed method was applied to blood samples from lymphoma patients, results consistent with the ELISA kit were obtained. This will open avenues for constructing multiple protein detection sensors.
Assuntos
Técnicas Biossensoriais , Nanotubos de Carbono , Humanos , Imunoensaio , Ligas , CalibragemRESUMO
Simultaneous quantification of multiple heavy metal ions (HMIs) is essential due to enhanced toxicity induced by synergistic effects. The currently available detection methods suffer from drawbacks such as costly devices, poor anti-interference, and specificity. In this work, a ratiometric electrochemical aptasensor for simultaneous detection of trace lead (Pb2+) and mercury ions (Hg2+) was developed. Here, a metal-organic framework, UiO-66-CNTs, with inherent and stable electrochemical signal was used for loading complementary strands (CSs) and internal reference. Guanine-rich and thymine-rich oligonucleotides, labelled with carbon dots (CDs), acted as aptamers (Apts) and hybridized with CSs to form M-shaped DNA complexes. Pb2+ and Hg2+ could be recognized and captured by Apts to form Pb2+-G-quadruplex and T-Hg2+-T complexes, leading to the destruction of M-shaped DNA complexes and changes in CDs' signal. The current ratios, ICDs/IUiO-66-CNTs, were applied to quantify Pb2+ and Hg2+. Benefiting from the anti-interference ability of the ratiometric strategy and the specificity of Apts, the proposed method enabled detection Pb2+ and Hg2+ through simple instrumentation, with detection limits of 2.0 ng mL-1 and 0.5 ng mL-1. Moreover, it was applied to assess Pb2+ and Hg2+ contamination in series of aquatic edible vegetables. The experimental results were consistent with inductively coupled plasma-mass spectrometry (ICP-MS), demonstrating its potential in practical applications.