This technology is a thrombin detection method that immobilizes fibrinogen on a gold electrode surface modified with a mercaptoundecanol linker, forms a fibrin monomer layer using thrombin, and then reacts the sample with ferrocene-labeled fibrinogen (Fc-Fib) to measure the aggregation amount based on thrombin enzymatic activity via redox current.
Immunoassays using hirudin peptides have faced challenges in accurate quantification because hirudin cannot bind to thrombin if it is already bound to other proteins. Methods involving the immobilization of aptamers on surfaces have also been limited by reduced protein activity due to aptamer binding, which decreases the accuracy of enzyme-based detection.
This technology selects mercaptoundecanol as the linker—which most efficiently forms the amount and orientation of fibrinogen assemblies among various alkanethiolates—and allows Fc-Fib to bind onto the fibrin monomer to obtain an electrochemical signal proportional to thrombin activity. Measured via differential pulse voltammetry, it demonstrates high selectivity and specificity across a wide concentration range from 1 ng/ml to 1 μg/ml. This method can be applied to electrochemical biosensors for diagnosing blood coagulation disorders and testing for lung metastasis tumor markers, enabling precise, low-cost, and compact quantification of thrombin without the need for antibodies or aptamers.
This invention was developed with support from the Ministry of Education, Science and Technology for the development of electrochemical biosensors utilizing bacteriophages.
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