Development of Functional Biointerface by Mixed Oligomeric Silatranes

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陳氏英紅(Tran Thi Anh Hong) 查詢紙本館藏

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生醫科學與工程學系

論文名稱

(Development of Functional Biointerface by Mixed Oligomeric Silatranes)

相關論文

★ 建立雙離子高分子修飾蛋白質技術與分析	★ DEVELOPMENT AND APPLICATIONS OF CATECHOL-FUNCTIONALIZED ZWITTERIONIC POLYMER
★ 雙離子矽氧烷共聚物以沉積法對聚二甲基矽氧烷進行生物相容性修飾

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摘要(中)

如今，生物感測器隨著醫療、病原體檢測、環境監測、食品安全檢測等廣泛應用而變得越來越重要。然而，這些樣品的成分相當複雜，會產生嚴重的背景信號導致，生物感測器受到影響。同時，在“非特異性吸附”造成降低靈敏度和特異性造成嚴重的問題。因此，通過低聚物氮矽三環(Oligomeric silatrans)對生物感測器進行表面修飾是改善這些問題的常用方法。巰基氮矽三環 (MPS) 是一種包含三環籠狀結構和跨環 N→Si 配鍵的化合物，提供可被控制的矽烷化並得到更均勻的薄膜。在本研究中，2-甲基丙烯醯氧基乙基磷醯膽鹼（MPC）或羧化聚乙二醇甲基丙烯酸酯（Carboxylated-PEGMA）將通過硫醇-烯加成反應與MPS進行化學鍵結，形成兩種低聚物氮矽三環MPS-MPCn和MPS-Carboxylated PEGMAm其具有各自獨立的功能。除此之外，MPC為雙離子材料，具有優異的抗汙性和良好的生物相容性。因此，在MPS-MPC2.5修飾後進行抗汙測試表現出優異的細菌粘附和蛋白質吸附的抵抗力。此外，低聚物 MPS-Carboxylated PEGMA2.5 作為大分子包含大量的羧基，通過共價鍵轉化為可功能化的中間體使其具有生物選擇性的表面。在生物檢測研究中，通過將 70% MPS-Carboxylated PEGMA2.5 和 30% MPS-MPC2.5 按濃度比例混合進行表面修飾，可以顯著減少非特異性吸附，並通過酵素結合免疫吸附試驗(ELISA)形成傳感表面。在大範圍的濃度測試中，混合的自組裝塗層表面在 25 µg/mL 至 1000 µg/mL 的多種抗原濃度範圍內展現出牛血清白蛋白(BSA)濃度與檢測信號之間的非線性關係，同時表現出與表面活性位點相連的生物受體從 5.5 µg/mL 到 27.5 µg/mL 的 HRP 偶聯二級抗體檢測的線性關係。因此，混合低聚物氮矽三環單一塗層是一種潛在的生物感測器表面改質方法，因其優秀的抗汙性能，可通過稀釋表面分子密度來提高目標檢測能力

摘要(英)

Nowadays, biosensor becomes more and more important along with a wide range of applications, including medical, pathogen discovery, environmental monitoring, food quality testing, etc. However, the components of these samples are quite complex and generate a serious background signal at the interface of biosensors. In addition, reducing sensitivity and specificity through non-specific adsorption of the biosensor are also a serious issue. Therefore, surface modification for biosensors via oligomeric silatranes has been developed to address these problems. Mercaptopropylsilatrane (MPS) is one type of silatranes containing the tricyclic caged structure and transannular N→Si dative bond that enables control silanization and provides thin homogeneous films. In this study, 2-methacryloyloxyethyl phosphorylcholine (MPC) and carboxylated poly(ethylene glycol) methacrylate (Carboxylated-PEGMA) can chemically linked with MPS through thiol-ene polymerization to form two kinds of copolymer MPS-MPCn and MPS-Carboxylated PEGMAm with independent functions. MPC is known as zwitterionic materials with excellent antifouling and good biocompatibility properties. Oligomeric MPS-MPC2.5 shows the excellent resistance of both bacterial adhesion and protein adsorption after modification. Moreover, oligomeric MPS-Carboxylated PEGMA2.5 as a bulky structure contains large carboxyl groups transforming into a functional intermediated product for bio-recognition elements by amine coupling chemistry. In the target detection study, surface modification by mixing 70% MPS-Carboxylated PEGMA2.5 and 30% MPS-MPC2.5 in the molar ratio could reduce significantly non-specific adsorption and develop a sensing surface in the enzyme-linked immunosorbent assay (ELISA). In a wide range of concentration tests, mixed oligomeric coatings demonstrated a non-linear response between bovine serum albumin concentration and detection signal in the multiple antigen concentrations from 25 µg/mL to 1000 µg/mL. In addition, it provided a linear response from 5.5 µg/mL to 27.5 µg/mL of HRP conjugated secondary antibody detection through a bioreceptor connected with an active site on the surface. Therefore, mixed oligomeric silatranes coatings are a potential approach to establish the interfacial system for biosensors because of the antifouling properties, improving target detection capability by increasing carboxyl group molecule surface’s density.

關鍵字(中)

★ 自組裝單層
★ 非特異性吸附
★ 雙離子材料
★ 抗汙性能
★ 2-甲基丙烯酰氧乙基磷酰胆碱 (MPC)
★ 巯基丙基硅油烷 (MPS)

關鍵字(英)

★ Self-assembled monolayer
★ nonspecific adsorption
★ zwitterionic materials
★ antifouling properties
★ 2-methacryloyloxyethyl phosphorylcholine (MPC)
★ mercaptopropylsilatrane (MPS).

論文目次

CHINESE ABSTRACT i
ABSTRACT iii
LIST OF FIGURES viii
LIST OF TABLES xii
LIST OF ABBREVIATIONS xiii
Chapter I: INTRODUCTION 1
1-1 BIOSENSOR 1
1-1-1 Classification base on the biological signal of biosensor 2
1-1-2 Factors affecting the signal of biosensor 3
1-2 Self-assembled monolayer (SAMs) 5
1-2-1 Silane functional group 6
1-2-2 Silatrane functional group 11
1-2-3 Mixed self-assembled monolayer (Mixed SAMs) 17
1-3 Antifouling materials 19
1-3-1 Nonspecific adsorption 19
1-3-2 Polyethylene glycol material 20
1-3-3 Zwitterionic materials 21
1-4 Thiol-ene polymerization 24
1-5 Methods of protein immobilization 25
1-6 Bovine serum albumin (BSA) immobilization on the surface 27
CHAPTER II: RESEARCH OBJECTIVES 29
Chapter III. MATERIALS AND METHOD 30
3-1 MATERIALS 30
3-1-1 Chemicals 30
3-1-2 Equipments 30
3-2 SYNTHESIS 31
3-2-1 Mercaptonitrosilyl tricyclic (MPS) 31
3-2-2 Carboxylated Poly(ethylene glycol) methacrylate 31
3-2-3 Copolymer MPS-MPCn and MPS-Carboxylated PEGMAm 32
3-3 EXPERIMENTAL METHOD 33
3-3-1 Pre-treatment of substrate 33
3-3-2 Preparation of self-assembled monolayer 34
3-3-3 Water contact angle measurement 34
3-3-4 Attenuated Total Reflectance-Fourier Transform Infrared spectrum (ATR-FTIR) measurement 35
3-3-5 Ellipsometer measurement 35
3-3-6 X-ray photoelectron spectroscopy (XPS) measurement 35
3-3-7 Hydrogen-1 Nuclear Magnetic Resonance Spectrometer (1H NMR) 35
3-3-8 Bacteria adhesion test 35
3-3-9 Protein adsorption test (Antifouling property) 36
3-3-10 Target detection ability 36
CHAPTER IV: RESULTS AND DISCUSSION 39
4-1 Investigating the antifouling properties of different chain lengths of MPS-MPCn modified silicon wafer 39
4-1-1 1H NMR spectrum of monomer MPS 39
4-1-2 1H NMR spectrum of oligomeric silatranes MPS-MPCn 40
4-1-3 Thickness measurement of different ratio MPS-MPCn 42
4-1-4 Contact angle measurement of MPS-MPCn 43
4-1-5 Protein adsorption test of MPS-MPCn modified sample 45
4-1-6 Bacterial adhesion test on MPS-MPCn modified sample 46
4-2 Biomolecule detection assay of mixing MPS-MPC2.5 and MPS Carboxylated PEGMAm 49
4-2-1 NMR spectrum of Carboxylated PEGMA 49
4-2-2 ATR-FTIR spectrum analysis of Carboxylated PEGMA and PEGMAOH 51
4-2-3 1H NMR spectra of MPS-carboxylated PEGMAm 53
4-2-4 XPS measurement of single oligomeric coating and mixed oligomeric coating after modified substrate 55
4-2-5 Contact angle measurement of MPS-carboxylated PEGMAm and mixed oligomeric coatings 57
4-2-6 Thickness measurement of MPS-carboxylated PEGMAm and mixed oligomeric coatings 58
4-2-7 The influence of different pH on BSA immobilization using ELISA) 59
4-2-8 The capability of BSA immobilization of single coatings and mixed coatings using ELISA 61
4-2-9 Exploring the influence of different concentration of BSA target on detection signal using ELISA 63
4-2-10 The ability of antibody detection of active site Carboxylated PEGMA 64
4-2-11 Investigating the relationship between the different concentration of antibody on detection signal (ELISA) 67
CHAPTER 5: CONCLUSION 68
CHAPTER 6: FUTURE PERSPECTIVES 69
REFERENCE 70

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指導教授

黃俊仁李宇翔(Chun-Jen Huang Yu-Hsiang Lee)

審核日期

2021-8-9

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