Ratiometric electrochemical immunoassay based on 2D Co/Fe MOF decorated with toluidine blue and Fc-labeled Schiff base for accurate assay of alpha-fetoprotein in clinical serum.

The high level of alpha-fetoprotein (AFP) expression is closely related to hepatocellular carcinoma (HCC). Herein, a dual signal ratiometric electrochemical immunosensor based on chitosan-ferrocenecarboxaldehyde-spindle gold (Chit-Fc-SAu) and Co/Fe metal-organic framework-toluidine blue/polydopamine (Co/Fe MOF-TB/PDA) was proposed for quantitative analysis of AFP. Specifically, Chit-Fc-SAu worked as a substrate to trap more primary antibodies (Ab1) generating the first electrochemical signal from Fc. Thanks to the large specific surface area, the synergistic and electronic effects of Co/Fe MOF nanosheets, and the rich functional groups of PDA, Co/Fe MOF-TB/PDA could load more secondary antibodies (Ab2) and signal molecules (TB) providing another amplified electrochemical signal. In the presence of AFP, Ab1-AFP-Ab2 formed a sandwich structure, and as the AFP concentration increased, the peak current ratio of TB to Fc (ITB/IFc) also increased. The dual signal ratiometric strategy can avoid environmental signal interference and achieve signal self-calibration, thereby improving the accuracy and reproducibility of detection. After a series of exploration, this self-calibrated ratiometric immunosensor exhibited a wide linear range (0.001-200 ng mL-1), a low detection limit (0.34 pg mL-1), and good repeatability. When applied to the assay of clinical serum samples, the detection results of ratiometric sensor were consistent with that of commercial electrochemiluminescence (ECL) immunoassay, significantly superior to that of non-ratiometric sensor. The self-calibrated strategy based on ratiometric sensor helps to improve the accuracy of AFP in clinical diagnosis.

Facile preparation of Ru nanoassemblies for electrochemical immunoassay of carcinoembryonic antigen in clinical serum.

Abnormal expression of carcinoembryonic antigen (CEA) can be used for early diagnosis of various cancers (e.g. colorectal cancer, cervical carcinomas, and breast cancer). In this work, using l-cysteine-ferrocene-ruthenium nanocomposites (L-Cys-Fc-Ru) to immobilize secondary antibody (Ab2) and Au nanoparticles (NPs) as the substrate to ensure accurate capture of primary antibody (Ab1), a signal-on sandwich-like biosensor was constructed in the presence of CEA. Specifically, Ru nanoassemblies (NAs) were first prepared by a facile one-step solvothermal approach as signal amplifiers for the electrical signal of Fc. Based on specific immune recognition, as the increase of CEA concentration, the content of L-Cys-Fc-Ru-Ab2 captured on the electrode surface also increased, thus the signal of Fc gradually increased. Therefore, the quantitative detection of CEA can be realized according to the peak current of Fc. After a series of experiments, it was found that the biosensor has a wide detection range from 1.0 pg mL-1 to 100.0 ng mL-1 and a low detection limit down to 0.5 pg mL-1, as well as good selectivity, repeatability and stability. Furthermore, satisfactory results were also obtained for the determination of CEA in serums, which were comparable to commercial electrochemiluminescence (ECL) method. The developed biosensor shows great potential in clinical applications.

Simple synthesis of PtRu nanoassemblies as signal amplifiers for electrochemical immunoassay of carbohydrate antigen 19-9.

In clinical analysis, carbohydrate antigen 19-9 (CA199) is a gold standard for pancreatic cancer diagnosis. Herein, PtRu nanoassemblies (NAs) were synthesized via a facile one-step solvothermal approach, with the help of octylphenoxypolye thoxyethanol (NP-40) acted as a growth-directing molecule, and triethylene glycol (TEG) worked as a reductant and solvent. During the assembly process of small particles, a large number of voids were formed, which significantly increase the specific surface area of the PtRu NAs exhibiting excellent electrocatalytic performance. Incorporating the PtRu NAs as signal amplifiers for potassium ferrocyanide oxidation into the specific molecular recognition of proteins, a facile signal-enhanced electrochemical (EC) immunosensor was developed. Verified by a series of experiments, the proposed immunosensor presented a wide linear range (10-4-70 U mL-1) and a low detection limit (3.3 × 10-5 U mL-1), accompanied by good reproducibility, selectivity, and stability, which could be applied in human serum samples for the determination of CA199, and was comparable to commercial electrochemiluminescence (ECL) immunoassay. Feasibility of batch fabrication of PtRu NAs makes nanomaterial-based EC immunoassay promising for the determination of similar cancer markers in future.