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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37717完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 劉子銘(Tzu-Ming Liu) | |
| dc.contributor.author | Fu-Lien Huang | en |
| dc.contributor.author | 黃甫簾 | zh_TW |
| dc.date.accessioned | 2021-06-13T15:40:11Z | - |
| dc.date.available | 2016-08-15 | |
| dc.date.copyright | 2011-08-15 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-10 | |
| dc.identifier.citation | References:
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37717 | - |
| dc.description.abstract | 紫質(Protoporphyrins)在身體代謝中扮演重要的角色，積累在腫瘤組織中的紫質為癌症的生物指標。然而，紫質在人體的癌症組織或細胞中含量很低，傳統偵測紫質方法不具足夠靈敏度以分析紫質。在此論文中，我們發展一個結合功能化磁性奈米粒子和質譜技術的新策略，能夠同時萃取和偵測紫質。
我們合成表面裝載三甘胺酸的功能化磁性奈米粒子，然後敖合鐵金屬離子，藉由鐵金屬離子和紫質中四個吡咯（pyrrole）環上的氮原子之間的親和力作用作為純化工具。使用紫質(protoporphyrins)標準品為模型，我們測試了奈米粒子在水溶液中萃取紫質的最佳化條件。測試結果顯示，使用60μg的奈米粒子能夠萃取0.1 nmole 紫質。奈米粒子萃取方法條件最佳化後，結合基質輔助雷射游離脫附質譜法(MALDI-TOF MS)能夠偵測到水溶液中紫質的(protoporphyrins)偵測極限（LOD）為10 nM。相對於傳統的高效液相層析法和螢光技術，奈米粒子能夠快速萃取紫質。為了評估此方法能夠在複雜的樣品中萃取紫質，我們挑選了有自然發生存在紫質的大腸細胞（CT26），並且加入標準品紫質，目前提供的檢測靈敏性為500nM 的紫質。因此，利用磁性奈米粒子結合質譜技術的靈敏性與快速性，本論文發展的新技術可應用於這些紫質的快速分析。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-13T15:40:11Z (GMT). No. of bitstreams: 1 ntu-100-R98548032-1.pdf: 4990831 bytes, checksum: feb97cf9354d0c8b1eb1f8b46c3deb4d (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 口試委員會審定書...........................................I
致謝..................................................... II 中文摘要..................................................IV Abstract ..................................................V Table of Contents .......................................VII List of Figures............................................X List of Tables .........................................XIII Abbreviation.............................................XIV CHAPTER 1 INTRODUCTION ................................... 1 1-1 Chemistry of Porphyrins............................... 1 1-1-1 Chemical structure of Porphyrins ....................1 1-1-1Biochemistry of Porphyrins .......................... 2 1-2 Clinical Aspects of Porphyrins.........................3 1-3Conventional Method for Porphyrins Analysis............ 5 1-3-1 Liquid Chromatography Analysis of Porphyrins........ 5 1-3-2 Fluorescence Analysis of Porphyrins................. 6 1-4 Thesis Motivation and Objectives...................... 7 CHAPTER 2 PRINCIPLES...................................... 9 2-1 Principle of Mass Spectrometry........................ 9 2-1-1 Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry ............. ...............................10 2-2 Extraction of Porphyrins by Magnet Metal Immobilized Nanoparticle .............................................12 2-2-1 Immobilized Metal Affinity Chromatography ..........12 2-2-2 Magnetic Nano-Materials for Puification of Phosphopeptides ......................................... 13 2-2-3 NTA-PEG@MNP for Enrichment of Porphyrin ........... 14 CHAPTER 3 MATERIALS AND METHODS.......................... 16 3-1 Fabrication of NTA-PEG@MNP............................16 3-1-1 Synthesis of Fe3O4@MNP............................. 16 3-1-2 Surface Blocking by PEG.............................16 3-1-3 Preparation of NTA-PEG@MNP. ....................... 17 3-2 Cell Culture......................................... 17 3-3 Cell Lysis. ......................................... 18 3-4 MALDI-TOF MS Analysis................................ 18 3-5 Enrichment of Protoporphyrin IX Using Fe3+-NTA-PEG@MNP.................................................. 19 3-5-1 General Protocol of Extraction Protoporphyrin IX... 19 3-5-2 Optimization of Extraction Protocol................ 20 3-5-2-1 Survey Suitable Metal Ion for the Binding on NTA-PEG@MNP ................................................. 20 3-5-2-2 pH Effects on Binding............................ 21 3-5-2-3 Effects of Ionic Medium on Binding............... 21 3-5-2-4 Reaction Time for Binding........................ 22 3-5-2-5 Targeting Capacity of NTA-PEG@MNP on PpIX ....... 22 3-5-2-6 Enrichment Efficiency of Fe3+-NTA-PEG@MNPs in Different Concentration.................................. 22 3-5-3 Optimization of MS Detection Condition............. 23 3-5-3-1 Invesitgate On-particle vs Elution with EDTA..... 23 3-5-3-2 Evaluate pH Effect on MS Detection............... 24 3-5-3-3 Effect of Elution Buffers for Facile PpIX Release.................................................. 25 3-5-3-4 Effect of Elution Buffer Concentration on Facile PpIX Release ............................................ 25 CHAPTER 4 Results and Discussion......................... 27 4-1 Fabrication and Characterization of NTA-PEG@MNP...... 27 4-2 Magnetic NTA-based Affinity Isolation for the Quantification of Porphyrins..............................29 4-2-1 Improved Detection of PpIX using Fe3+-activated NTA-PEG@MNP.................................................. 31 4-2-2 Optimization of Charging of Metal Ions on NTA-PEG@MNP.................................................. 32 4-2-2-1 Activation of NTA-PEG@MNP with Transition Metals. 33 4-2-2-2 Effect of pH on Binding of PpIX with Fe3+-activated NTA-PEG@MNP.............................................. 35 4-2-2-3 Effect of Ionic Medium........................... 37 4-2-2-4 Effect of Incubation Time........................ 39 4-2-2-5 Capacity of NTA-PEG@MNP.......................... 41 4-3 Optimization of Detection and Ionization of Protoporphyrin IX by NTA-PEG@MNP and MALDI-TOF MS.........43 4-3-2 Optimization of Elution of PpIX by Competition Molecule ................................................ 44 4-4 Concentration Effect................................. 47 4-5 Sensitivity of On-particle Detection with Fe3+-activated NTA-PEG@MNP.....................................48 4-6 Extraction and Detection of PpIX from CT26 Colorectal Cancer Cell.............................................. 50 CHAPTER 5 CONCLUSION..................................... 54 References: ............................................. 56 | |
| dc.language.iso | en | |
| dc.subject | 質譜 | zh_TW |
| dc.subject | 磁性奈米粒子 | zh_TW |
| dc.subject | 紫質 | zh_TW |
| dc.subject | porphyrin | en |
| dc.subject | MALDI-TOF MS | en |
| dc.subject | Magnetic Nanoparticle | en |
| dc.title | 發展功能化磁性奈米粒子結合質譜技術偵測內生性紫質 | zh_TW |
| dc.title | Magnetic NTA -Based Affinity Mass Spectrometry for Rapid Detection of Indigenous Porphyrins | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 陳玉如(Yu-Ju Chen) | |
| dc.contributor.oralexamcommittee | 周綠蘋(Lu-Ping Chow),張富雄(Fu-Hsiung Chang) | |
| dc.subject.keyword | 質譜,磁性奈米粒子,紫質, | zh_TW |
| dc.subject.keyword | MALDI-TOF MS,Magnetic Nanoparticle,porphyrin, | en |
| dc.relation.page | 62 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-10 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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