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Current advances in molecular imprinting has many drawbacks such as the lack of specificity, this is the inability to selectively detect one single species of biomolecule. In addition, it is very difficult to produce a device that accounts with high sensitivity to detect relevant biomolecules and generate reproducible data from complex samples. The present project aims to offer an alternative by developing a technique to synthesize a polymeric matrix with molecularly imprinted cavities that excludes non-targeted species and binds to the targeted molecule, this synthetic platform in combination with proven efficient surface plasmon resonance technology achieve high sensitivity, for real time monitoring of molecular binding.Polyacrylamide is a hydrophilic and biocompatible synthetic polymer used in industry and in the lab. It is used in chromatography, electrophoresis, and recently it has been used in molecular imprinting. However conventional techniques for molecular imprinting using acrylamide as the functional monomer are not yet robust enough due to the high swelling ratio in the presence of water. In the present study, molecularly imprinted polyacrylamide was produced in nano-thin films on top of the gold surface of a surface plasmon resonance sensor. The model template used in this study was lysozyme and bovine serum albumin because they are relatively big and complex proteins that easily solubilize in water. The system was evaluated by comparing signals when exposing the imprinted polyacrylamide to a solution containing lysozyme. Uv-Vis spectrophotometry, scanning electron microscopy (SEM), matrix assisted laser desorption ionization coupled with time of flying mass spectrometry (MALDI-TOF MS), and surface plasmon resonance (SPR) were used to characterize the novel biosensor. The results obtained from this project showed that it is possible to prepare a molecularly imprinted system with high sensitivity and selectivity that is stable and offers reproducibility in the detection of proteins. |
