Development of a functional impedimetric immunosensor for accurate detection of thyroid-stimulating hormone.

Thyroid-stimulating hormone (TSH), which regulates the synthesis of thyroid gland hormones affecting the whole metabolism, is a pituitary hormone. Determination of TSH is crucial for monitoring thyroid gland-related disorders and some metabolic diseases.In this study, a nonlabeled immunosensor based on covalent immobilization of anti-TSH antibody by using the formation of self-assembled monolayers (SAM) of 4-mercaptophenylacetic acid (4-MPA) and functionalization of carboxyl ends with 1-ethyl-3-(3-dimetilaminopropil) carbodiimide (EDC)/N-Hydroxysuccinimide (NHS) was fabricated for detection of TSH. Immobilization steps including the concentration of 4-MPA, the concentration of anti-TSH antibody, and duration of anti-TSH antibody incubation were optimized by utilizing electrochemical impedance spectroscopy. Under optimal conditions, a sensitive, rapid, and accurate determination of TSH at a concentration range between 0.7 and 3.5 mIU/L was accomplished with a notable linearity and LOD value of 0.034 mIU/L, as well as reproducibility and repeatability. Moreover, for comparison, linear range experiments were also carried out by using other electrochemical methods, including linear sweep voltammetry, cyclic voltammetry, and capacitance spectroscopy. Finally, the constructed immunosensor was used for analyzing TSH levels spiked in the artificial serum samples.