Preparation of Molecular Imprinting Based Lysozyme Sensors

Abstract

Detection of proteins has strategic importance due to their possible usability as disease biomarkers, environmental monitoring, food quality monitoring, and çivil defence purposes. For the detection of proteins there are many methods and devices but they all have some drawbacks like low sensitivity and high detection limits. Detection of proteins has strategic importance due to their possible usability as disease biomarkers, environmental monitoring, food quality monitoring, and çivil defence purposes. For the detection of proteins there are many methods and devices but they all have some drawbacks like low sensitivity and high detection limits. This study aims to produce molecular imprinting based polymeric biomimetic receptors that is bound to the surface of the gold coated chips of the Surface Plasmon Resonance (SPR) system by exploiting the unique distribution of charged, hydrophobic and hydrophilic groups on the surface of every protein, in this case lysozyme. Lysozyme is a protein with a well-known primary structure. Due to its small size and simple molecular structure, lysozyme has been frequently chosen as a unique model protein in developing of new detection methods. Artificial receptors for lysozyme prepared by micro-contact imprinting of lysozyme to the surface initiated polymeric ultra-thin films. By this method complementary cavities for lysozyme obtained on the surface initiated polymeric ultra-thin film. To increase recognition capability of the sensors positive, negative and hydrophobic side chained polymerizable amino acids, N-methacryloyl L-histidine (MAH), N-Methacryloyl-L-aspartic acid (MAAsp) and N-methacryloyl L-tryptophan (MATrp), used together during imprinting process. According to the data obtained from modeling analysis of Lysozyme functional monomers cross-linkers selected and also proper functional monomer ratio estimated according to this data. Prepared SPR sensor used for the detection of lysozyme in variety of body fluids like saliva, mucus, tear and urine. Also force-distance measurements of imprinted, non-imprinted and bare sensor obtained by Lysozyme bound Atomic Force Microscopy tips. According to the force distance measurements imprinted surface have 1,3 times higher affinity than the non-imprinted sensors