Ammonia-trapping multilayer polymer urease film amplifier coated surface plasmon resonance sensor for ultra-sensitive urea detection

Abstract

In this paper, we have developed a new urea signal amplification technology based on a multilayer urease polymer film self-assembly structure for the first time, enabling the realization of ultra-accurate urea optical detection. We demonstrate the effectiveness of this technique based on surface plasmon resonance (SPR) platform. When the urea signal amplifier is combined with SPR sensors, multilayer urease polymer film contacts with urea molecules, the urea molecules are oxidated into NH3 and CO2, causing the pH increase, which weakens the electrostatic interaction between different polymer layers and induces the RI reduction of the multilayer urease film, thereby causing the shift of the SPR spectrum. Experiment results confirm that urea sensitivity has been improved 12-fold, and the limit of detection (LOD) has been optimized to less than 10−3 mmol/L. Furthermore, the anti-interference has been improved 22.8-fold and is kept even the interference signal strength increases by 100 times. Moreover, multilayered urease polymer leads to the increase of urease on the sensor surface, thus expanding the detection range of urea by 3.5 times. This provides a new way to solve the contradiction between high sensitivity and large measurement range of optical fiber biosensor. Importantly, the concept of using enzyme-loaded polymer films for signal amplification can be adapted for the detection of various other biomolecules by replacing the enzyme with a specific recognition element for the target molecule. This opens doors for developing highly sensitive and specific biosensors for diverse applications in healthcare, environmental monitoring, and food safety.