In Vivo Detection and Absolute Quantification of a Secreted Bacterial Factor from Skin Using Molecularly Imprinted Polymers in a Surface Plasmon Resonance Biosensor for Improved Diagnostic Abilities

In this study, a surface plasmon resonance (SPR) biosensor was developed for the detection and quantification of a secreted bacterial factor (RoxP) from skin. A molecular imprinting method was used for the preparation of sensor chips and five different monomer-cross-linker compositions were evaluated for sensitivity, selectivity, affinity, and kinetic measurements. The most promising molecularly imprinted polymer (MIP) was characterized by using scanning electron microscopy, atomic force microscopy, and cyclic voltammetry. Limit of detection (LOD) value was calculated as 0.23 nM with an affinity constant of 3.3 X 10(-9) M for the promising MIP. Besides being highly sensitive, the developed system was also very selective for the template protein RoxP, proven by the calculated selectivity coefficients. Finally, absolute concentrations of RoxP in several skin swabs were analyzed by using the developed MIP-SPR biosensor and compared to a competitive ELISA. Consequently, the developed system offers a very efficient tool for the detection and quantification of RoxP as an early indicator for some oxidative skin diseases especially when they are present in low-abundance levels (e.g., skin samples).