溶菌酶(lysozyme)為一可診斷多種病徵之生物指標，故臨床檢驗血清、尿液或唾液中的溶菌酶之濃度可協助醫生判斷病人的疾病，一般在臨床醫事檢驗上，常以免疫分析法來檢測，但在臨床上應用之免疫分析方法，不但步驟繁雜，且因自然界的抗原、抗體不易取得，所以分析成本昂貴。分子模版因具有與天然抗體相似之專一性，並且穩定性佳、製備簡單且成本低廉之諸多優點，若引用分子模版取代天然抗體，則可解決免疫分析時所需高價或稀有之天然抗體的缺點。

　　本研究應用新穎的微接觸技術於製備溶菌酶分子模版，此方法的優點在於不需考慮目標分子對於高分子單體或其他溶劑的溶解度問題。在此研究中，主要是應用酵素連結免疫吸附分析法(ELISA)搭配冷螢光儀或掃瞄式電化學顯微鏡(SECM)為分析元件以鑑定溶菌酶分子模版與非分子模版之吸附效能，且可由掃瞄式電化學顯微鏡在操作時所得到之電流變化訊號可繪製出被掃瞄物件之表面輪廓圖。並分別探討交聯劑與功能性單體對分子模版吸附效能之影響。結果發現在不同交聯劑時，使用TEGDMA製備分子模版會比其他交聯劑得到較佳吸附效能；如果混合不同功能性單體，結果發現當高分子混合溶液為TEGDMA：styrene monomer: HEMA: acrylamide以莫耳比為10:1:1:1時所製備的分子模版會有最佳的吸附力，它比使用單一高分子單體所製備出的分子模版之吸附效能更高。將此分子模版置於0.01 mg/mL溶菌酶溶液中吸附2小時後可得到最大吸附效能。此外，並應用SEM及AFM等顯微鏡技術來檢視分子模版表面，由所得到之輪廓圖可觀察得經由聚合反應所形成之辨識位置；在分子模版的選擇性方面，無論分子模版於單成份、雙成份或三成份的蛋白質混合溶液中進行再吸附實驗，都具有辨識原目標物(溶菌酶)之能力。

　　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.

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