Aptamer-free electrochemical detection of thrombin based on coagulation reaction of ferrocene-labeled fibrinogen

• A new electrochemical method for aptamer-free thrombin detection has been developed.
• Fibrinogen was covalently coupled with a ferrocene as an electroactive label.
• Coagulation of ferrocene-labeled fibrinogen produces electrochemical signal.
• The method showed high specificity for only thrombin among various proteins.

A new electrochemical method for aptamer-free thrombin detection has been developed by using a fibrinogen-modified electrode and the coagulation reaction of fibrinogen labeled with ferrocene (Fibno-Fc) by enzymatic reaction of thrombin. Fibrinogen was covalently coupled with a ferrocene as an electrochemically active label. The fibrinogen-modified electrode was immersed in a mixed solution of Fibno-Fc and target thrombin. Thrombin catalyzes a cleavage reaction of Fibno-Fc resulting in the coagulation on the modified electrode surface. The surface adsorbed Fc moiety produced voltammetric signal for ferrocene oxidation which is proportional to the concentration of target thrombin with a good linear relationship in the range of 100 pg/ml to 1 μg/ml. The method showed high specificity for only thrombin among various proteins.

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