應用新穎微接觸技術製備溶菌酶分子模版之研究

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應用新穎微接觸技術製備溶菌酶分子模版之研究

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

陳筱淇 , Masters　　Advisor：許梅娟;周澤川　　

繁體中文

掃瞄式電化學顯微鏡；酵素連結免疫吸附分析；分子模版；溶菌酶；微接觸技術； SECM ； microcontact printing ； Lysozyme ； molecularly imprinted polymer ； ELISA

Abstract │ Reference (64) │ Times Cited (5) │ Altmetrics

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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.

Reference ( 64 ) 〈TOP〉

[3] V.A. Proctor, F.E. Cunningham, "The chemistry of lysozyme and its use as a food preservative and a pharmaceutical," CRC Critical Reviews in Food Science and Nutrition, vol. 26, pp. 359, 1988.
連結：
[4] D.M. Rothwarf, H.A. Scheraga, "Role of non-native and hydrophobic interaction in the folding of hen egg white lysozyme," Biochemistry, vol. 35, pp. 13797, 1996.
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17349.htm.", 2005/07/14.
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[10] J. Chongqiu, L. Li, "Lysozyme enhanced europium-etacycline complex fluorescence: a new spectrofluorimetric method for the determination of lysozyme," Analytica Chimica Acta, vol. 511, pp. 11-16, 2004.
連結：
[11] K. Hirayama, Y. Sakai, K. Kameoka, "Synthesis of polymer particles with specific lysozyme recognition sites by a molecular imprinting technique," Journal of Applied Polymer Science, vol. 81, pp. 3378-3387, 2001.
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