Target detection kit containing a thrombin label

Visual Target Detection Kit Using Thrombin-Fibrinogen Coagulation

This technology is a target detection kit consisting of a thrombin-conjugated antibody, a biochip immobilized with target-specific antibodies or probes, and fibrinogen, a substrate for thrombin. When a target substance is captured on the biochip, the thrombin-labeled antibody binds to it, and the subsequently added fibrinogen coagulates into fibrin, indicating the presence of the target.

Existing detection methods utilizing blood coagulation mechanisms have been hindered by complex procedures and difficulties in achieving accurate measurements. It has also been challenging to intuitively verify the quantity of target substances without separate analytical equipment.

This technology links thrombin to an antibody via a thrombin aptamer and a streptavidin-antibody complex, allowing fibrin coagulation to proceed according to the amount of captured target substance. The coagulation process can be visually confirmed, and signals can be amplified by adding color particles, quantum dots, or gold particles. It can be applied to point-of-care diagnostic kits for detecting antigens, nucleic acids, metal ions, and glucose, and can be expanded into various diagnostic and analytical products simply by replacing the detection probe.

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Key Features:

• A target detection kit comprising a thrombin-conjugated antibody, a biochip with target-specific antibodies or probes immobilized on its surface, and fibrinogen
• A thrombin-conjugated antibody consisting of a streptavidin-antibody complex, a thrombin aptamer bound to the streptavidin, and thrombin bound to the aptamer
• A process of forming a thiol-terminated monolayer via silanization, immobilizing biotin using maleimide-PEO2-biotin, and then immobilizing thrombin onto the thrombin aptamer
• A process of sequentially reacting the target substance, thrombin-conjugated antibody, and fibrinogen on the biochip surface while mixing in signal-enhancing particles

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