發展快速篩選磷酸化胜□的方法：1.以新型金奈米表面電漿共振為感測方法 2.以第五族金屬氧化物磁性奈米粒子為親和萃取磷酸化胜□之探針

Abstract

金奈米粒子具有獨特的表面電漿共振光學特性，利用此特性，本論文的第一部分嘗試將金奈米粒子自組裝於透明玻璃片上，並在金奈米粒子上塗佈一層二氧化鈦薄膜，以二氧化鈦薄膜對於磷酸根的高親和性結合金奈米粒子的表面電漿共振特性，可簡單利用其在吸收光譜中最大吸收波長及最大吸收值的改變即可對樣品中的磷酸化胜□進行感測，達到快速分析磷酸化胜□的目的，實驗結果也使用MALDI-MS確認分析結果。本論文的第二部分也嘗試發展以包覆有氧化鈮及氧化鉭的第五族金屬氧化物磁性奈米粒子為磷酸化胜□之親和探針，發展新型的金屬氧化物親和層析法。此二種新型探針可針對複雜樣品中的微量磷酸化胜□進行選擇性萃取，再結合MALDI-MS分析進行確認，並配合蛋白質資料庫進行目標蛋白質的序列比對。此二種快速分析方法的發展應有利於目前的磷酸化蛋白質體的研究。

Gold nanoparticles have a unique optical property, i.e., surface plasmon resonance (SPR). On the basis of this feature, the first part of this thesis demonstrates a new sensing method for phosphopeptides by using titania-coated gold nanoparticles immobilized on the surface of a glass slide as the sensing substrate and using UV-visible absorption spectrophotometry as the detection tool. Because of the high affinity of titania for phosphopeptides, the detection principle is based on the shift of wavelength and intensity of optical absorption due to localized SPR (LSPR) of the gold nanoparticles induced by binding of phosphorylated species. The LSPR results are also confirmed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Additionally, the second part of this thesis shows new types of Group 5 metal oxide affinity chromatography (MOAC). That is, both niobium oxide and tantalum oxide coated magnetic beads have been explored to have the capability of interacting with phosphopeptides. These two new types of probes can be used as the affinity probes to selectively enrich phosphopeptides from complex samples. The phosphopeptides trapped on the affinity probes are then characterized by matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analysis followed by identification via protein database searching. The methods demonstrated herein should benefit the research in phosphoproteomics.