The Study of Fabricating Lysozyme-Imprinted Polymers by A Novel Microcontact Printing Method

• Main Authors: Hsiao-Chi Chen, 陳筱淇
• Other Authors: Mei-Jywan Syu
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/46679029348182554730

碩士 === 國立成功大學 === 化學工程學系碩博士班 === 93 === 　　Lysozyme is known as an important biological index for the diagnosis of various diseases. The lysozyme concentrations in serum, urine or spit are very important determinants to help doctors to diagnose a patient. In general, lysozyme can be detected by immunoassay in clinical assays. However, clinical immunoassays have some disadvantages, such as complication and high price of antibody and antigen in nature. The advantages of molecularly imprinting polymers (MIPs) are in terms of their superior stability, low cost and ease of preparation when MIPs compare to the production of natural receptors, antibodies and enzymes. Molecularly imprinted polymer is designed to replace the natural antibody, and it can also overcome the disadvantages of the stability of natural antibody in immunoassay. 　　In this study, the lysozyme-imprinted polymers were fabricated by a novel microcontact printing method. The benefit of the presented method was the solubility of template in the polymer solutions can be resolved. Using an enzyme-linked immunosorbent assay (ELISA) integrated with chemiluminesence (CL) or scanning electrochemical microscopy (SECM) as analytical element to measure rebinding efficiency of lysozyme to lysozyme-imprinted (Lys-MIP) and non-imprinted polymer (Non-MIP). And it could get the contour of lysozyme rebinding area on the lysozyme-imprinted polymer based on the different current by SECM. The effect of using different crosslinkers and functional monomers in the molecularly imprinted polymer was also investigated, individually. When using only single crosslinker as the monomer solution, the results indicated that the tetraethyleneglycol dimethacrylate (TEGDMA) based molecular imprinted polymer could get a highest rebinding efficiency than using other crosslinkers. We also found the highest performance of molecular imprinted polymer can be synthesized with the monomer solution composed TEGDMA: styrene: 2-hydrozyethyl methacrylate (HEMA): acrylamide with the mole ratio of 10:1:1:1. The maximum rebinding efficiency of lysozyme on the molecularly imprinted polymer was achieved in 0.01 mg/ml lysozyme solution and 2 hrs incubation. The surface morphologies of the molecularly imprinted polymer were monitored by a scanning electron microscope (SEM) and an atomic force microscope (AFM). The selectivity of the lysozyme-imprinted polymer showed very good recognition to lysozyme no matter in a single, binary or trinary protein solution.