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研究生:劉育如
研究生(外文):Liu, Yu-Ju
論文名稱:以EpCAM作為生物標記物發展新的循環腫瘤細胞檢測平台
論文名稱(外文):Development of a New Detection Platform for Circulating Tumor Cells with EpCAM as a Biomarker
指導教授:梁美智梁美智引用關係
指導教授(外文):Liang, Mei-Chih
口試委員:袁俊傑楊裕雄梁美智
口試委員(外文):Yuan, Chiun-JyeYang, Yuh-ShyongLiang, Mei-Chih
口試日期:2021-11-10
學位類別:碩士
校院名稱:國立陽明交通大學
系所名稱:生物科技學系
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2021
畢業學年度:110
語文別:英文
論文頁數:59
中文關鍵詞:循環腫瘤細胞檢測晶片癌症抗體
外文關鍵詞:Circulating Tumor Cellsensing chipcancerantibodyEpCAM
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  • 被引用被引用:0
  • 點閱點閱:109
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摘要 i
Abstract ii
Contents iii
List of abbreviations vi
Chapter 1. Introduction 1
1.1 Circulating Tumor Cell (CTC) 1
1.1.1 Introduction of circulating tumor cell 1
1.1.2 The advantage of circulating tumor cell detection 2
1.1.3 The common methods of circulating tumor cell detection 2
1.4 Epithelial cell adhesion molecule (EpCAM) 5
1.5 Introduction of Biosensors 5
1.6 The working principle of the CMOS 3D-NEA sensing chips 6
1.7 Electrochemical impedance spectroscopy (EIS) 7
Specific aims 8
Chapter 2. Materials and Methods 9
2.1 Materials 9
2.1.1 Chemicals 9
2.1.2 Buffers 10
2.1.3 Reagents 10
2.1.4 Antibodies 11
2.1.5 Equipment 11
2.1.6 Cell culture media and reagents 12
2.1.7 Miscellaneous items 12
2.1.8 The complementary metal oxide semiconductor (CMOS) 3D nano-electrode array sensing chip 13
2.2 Methods 13
2.2.1 Cell culture 13
2.2.2 Preparation of whole cell lysates 13
2.2.3 Protein quantification 14
2.2.4 Sodium Dodecyl Sulfate polyacrylamide gel (SDS-PAGE) preparation 14
2.2.5 Sample preparation 15
2.2.6 Western blot analysis 15
2.2.7 Immunofluorescence staining assay 16
2.2.8 immunocapture-based ELISA (IC-ELISA) 18
2.2.9 Surface modification and probe immobilization of chips 19
2.2.10 Blocking test for chips 21
2.2.11 The sensing chip with EpCAM antibody to detect cancer cells 22
2.2.12 Measurement of electrochemical impedance spectroscopy (EIS) 23
2.2.13 Statistical analysis 23
Chapter 3. Results 24
3.1 Comparison of the EpCAM expression levels in three cancer cells 24
3.2 Comparison of the EpCAM expression levels on the surface among three cancer cells 24
3.3 The cancer cells detection with EpCAM as biomarker by immunocapture-based ELISA. 25
3.4 The blocking influences the empty chip 26
3.5 The blocking influences the CMOS 3D-NEA sensing chip after surface modification 27
3.6 MCF-7 cancer cell lines detection by impedance measurement 27
3.7 The specificity and lower detection value of MCF-7 cancer cells by impedance measurement 28
Chapter 4. Discussion 30
Reference 34
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