Molecular imprinted nanoparticle assisted surface plasmon resonance biosensors for detection of thrombin

In this study, we designed a surface plasmon resonance (SPR) biosensors based on molecular imprinting to detect low amounts of thrombin directly in a short time, both in thrombin aqueous solutions and in patient serum samples. For this purpose, the thrombin imprinted and non-imprinted SPR biosensors were prepared by integrating the synthesized thrombin imprinted and non-imprinted nanoparticles on the allyl mercaptan modified gold SPR chip surface. The kinetic studies were performed with a thrombin concentration range from 0.1 to 400 pM with a detection limit of approximately 0.017 pM in thrombin aqueous solution. Morever, the limit of detection was found to be 0.033 pM in patient serum samples. When the selectivity of the thrombin imprinted SPR biosensors was compared with the competing molecules as bovine serum albumin and lysozyme, it was showed that the thrombin was 4.58 times and 3.99 times more selective than the bovine serum albumin and lysozyme molecules, respectively. The shelf life and reusability of the designed SPR biosensor showed that there was a 12.93% decrease in the performance according to the kinetic analyzes that performed after 8 months. In addition, thrombin detection from the patient serum samples was performed using both SPR biosensor and the enzyme-linked immunosorbent assay methods, and were calculated recoveries.