胜肽拓印聚乙烯乙烯醇分子模版於尿液中再生蛋白之萃取及感測

再生胰島蛋白1(Regenerating Islet-Derived Protein Family, Reg1)在人體為胰臟癌的重要指標分子，Reg1分為1α及1β，但是Reg1蛋白質非常昂貴，因此本研究選取Reg1表面胜肽之7個不同部分的胺基酸序列片段作為拓印目標分子，而以不同乙烯醇比例的聚乙烯乙醇為智慧材料，結合分子拓印技術製備其感測薄膜電極，選擇辨識效率最佳的胺基酸序列片段Peptide4, 7及32mol%乙烯醇比例之聚乙烯乙烯醇，以電化學方式檢測真實檢體中Reg1濃度。化學分析電子光譜儀(ESCA)和原子力顯微鏡(AFM)分析薄膜表面，清洗前後及再吸附氮含量的變化、表面粗糙度會因洗滌後而提高。藉由電化學感測尿液檢體中的Reg1，常人濃度低於5 ng/mL，病人濃度則高於60 ng/mL。進而製備拓印Peptide7之磁性奈米複合微粒，從大腸桿菌(E.coli)中萃取大腸桿菌代謝液中Reg1β，平均每次可萃取出11.66±1.69 pg/mL的Reg1，再以電子顯微鏡(TEM)及超導量子干涉儀(SQUID)等進行粒子型態及磁通量分析。

關鍵字

再生蛋白；尿液；分子拓印；聚乙烯乙烯醇；大腸桿菌

並列摘要

Regeneration of islet protein 1 (Regenerating Islet-Derived Protein Family, Reg1) is in the human body as an important indicator of pancreatic cancer. Reg1 divides into 1α and 1β, but Reg1 proteins is very expensive. So, in this study, we selected Reg1 surface peptides the seven different parts of the amino acid sequence fragment as rubbing the target molecule, and in different proportions of vinyl alcohol, poly(ethylene-co-vinyl alcohol) as smart materials, integrate molecularly imprinting technology which senses the film electrode preparation, choose to identify the most efficient amine amino acid sequence fragment Peptide4, 7 and 32mol% of vinyl alcohol, electrochemically detect the sensor really sample Reg1 concentration. Electron spectroscopy for chemical analysis (ESCA) and atomic force microscope (AFM) analysis of the film surface before and after washing and rebinding of nitrogen content, the surface roughness will increase after washing. By electrochemical sensing urine sample in Reg1, healthy concentration of less than 5 ng / mL, the pdac concentrations higher than 60 ng / mL. Further imprinted Peptide7 preparation of magnetic nanoparticles extraction from E. coli metabolic solution Reg1 (E.coli), the average can extract 11.66 ± 1.69 pg / mL of Reg1β, then to transmission electron microscopy(TEM) and superconducting quantum interference device(SQUID) patterns and the particle analysis.

並列關鍵字

Regenerating protein ； Urine ； Poly(ethylene-co-vinyl alcohol)(EVAL) ； Molecular imprinted polymers ； E.coli

參考文獻

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胜肽拓印聚乙烯乙烯醇分子模版於尿液中再生蛋白之萃取及感測

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