Development of a label-free immunosensor system for detecting oocyte maturation-inducing hormone in fish

A label-free immunosensor for detecting the oocyte maturation-inducing hormone 17, 20 beta-dihydroxy-4-pregnen-3-one (DHP), was developed. A principle of the sensor system was based on differences in electrochemical activity changed by an immunoreaction in the absence and presence of DHP. For preparation of the immunosensor, anti-DHP IgG was immobilized on an Au working electrode modified with a self-assembled monolayer of 3-mercaptopropionic acid. The sensor was immersed into a sample solution containing DHP. DHP was determined by cyclic voltammetry. The immunosensor showed a specific response to DHP, and the oxidation peak current linearly decreased in the range of 7.8-500 pg ml(-1) with a correlation coefficient of 0.996. The sensor system was applied to determine the DHP levels in plasma of goldfish and was compared with the DHP levels of the same samples determined using an ELISA as the conventional method. Good correlation was obtained between values determined using both methods in the range of 0.1-7.7 ng ml(-1) (correlation coefficient 0.876). These findings suggest that the proposed label-free immunosensor can be used to analyze DHP levels in fish plasma samples.