Carbohydrate antigen 199 (CA199) is the most sensitive marker reported for pancreatic cancer, and it is a difficult task to develop a highly sensitive assay for CA199. During the experiment, a ratiometric electrochemical immunosensor for quantitative analysis of CA199 was prepared for NiCo@Fc-MWCNTs-LDH as the electrode sensing surface and 3D-rGOF@Ag/Au as the label of Ab2. NiCo@Fc-MWCNTs-LDH not only provide the required signal for the immunosensor, but also have a layered structure to obtain a large specific surface area, which can provide more sites for the placement of biological molecules. rGOF has the advantages of large specific surface area and high porosity, which can adsorb Ag electrochemical probe through redox reaction. The modification of gold nanoparticles can not only enhance the electrical conductivity of nano-composites, but also immobilize more biomolecules to improve the sensitivity of electrochemical sensors. With the beefing up of CA199 concentration, the oxidation peak current of Ag increases and the oxidation peak current of Fc-COOH decreases. The ratio (y = IAg/IFc-COOH) of two different signals was linear with the logarithm of CA199 concentration in a certain value range. Under optimal conditions, the immunosensor showed excellent performance in the concentration range of 0.0001 U/mL to 10 U/mL, and the detection limit was 5.55 × 10-4 U/mL. The strategy could clearly discriminate between matched and mismatched targets, demonstrating high specifificity. This approach further detects CA199 in human plasma to differentiate pancreatic cancer patients from healthy individuals with high accuracy. This method also provided a new idea for the ultrasensitive quantitative detection of other biomarkers.
Keywords: Carbohydrate antigen199; Layered double hydroxides; Ratiometric electrochemical immunosensor; Reduction of graphene oxide.
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