Enzyme-loaded liposome with biocatalytic precipitation for potentiometric immunoassay of thyroid-stimulating hormone in thyroid carcinoma

摘要

A simple and feasible potentiometric immunosensing platform based on enzymatic biocatalytic precipitation technique was designed for the sensitive detection of thyroid-stimulating hormone(TSH;a typical kind of biomarkers for thyroid carcinoma),using horseradish peroxidase(HRP)-loaded liposome for the signal amplification.To construct such an assay system,a sandwich-type immunoreaction was readily carried out on monoclonal anti-TSH capture antibody-coated electrode by using polyclonal antiTSH secondary antibody-conjugated HRP-loaded liposome.Accompanying the formation of sandwichtype immunocomplex,the carried liposome was lysed through the added Triton X-100 to release the entrapped HRP molecules,which catalyzed the oxidation of 4-chloro-1-naphthol to produce an insoluble and uncharged organic precipitation on the electrode surface,thereby causing the change of the local electrical potential.Two labeling protocols with and without the liposome were investigated for detection of target TSH,improved analytical features were achieved with HRP-entrapped liposome.Under optimal conditions,the potentiometric immunosensor had good responses for TSH detection within the linear range of 0.01-30 p IU/mL at a detection limit of 0.0067 p IU/mL.Good reproducibility,high specificity and long-time stability were acquired during the assay procedure.Importantly,a wellmatched accuracy between the potentiometric immunosensor and commercial human TSH ELISA kit was gave for the analysis of human serum samples.