Novel Sol-Gel Nanoporous Materials, Nanocomposites and Their

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Novel Sol-Gel Nanoporous Materials, Nanocomposites and Their Novel Sol-Gel Nanoporous Materials, Nanocomposites and Their Applications in Bioscience A Thesis Submitted to the Faculty of Drexel University by Zhengfei Sun in partial fulfillment of the requirements for the degree of Doctor of Philosophy September 2005 © Copyright 2005 Zhengfei Sun. All Rights Reserved. 2 Dedications This dissertation is dedicated to my parents, Mr. Chongzhen Sun and Mrs. Xiuqing Nie for their encouragement, support and love. 3 Acknowledgments In retrospect as I approach the completion of my doctorate, I feel a deep gratitude towards many people for their assistance and support. I would like to express my genuine gratitude to each of them, although it would be impossible for me to name all. First of all, I would like to sincerely thank my advisor, Dr. Yen Wei, for his tremendous time and effort spent in leading, supporting and encouraging me during the last five years. His passion for challenges has given me inspiration; his knowledge of science has given me guidance; his perseverance in research has given me confidence. Without his help and effort, it would be impossible for me to even get close to this point. I want to express my gratitude to all the committee members in my candidacy examination and/or my dissertation defense, Dr. Anthony Addison, Dr. Jean-Claude Bradley, Dr. Joe Foley, Dr. Susan Jansen-Varnum, Dr. Caroline Schauer, Dr. Sally Solomon and Dr. Jian-Min Yuan for their time and valuable suggestions. Special thanks are due to the committee chair, Dr. Anthony W. Addison for insightful discussions on many topics, including my oral proposal and research. I am also grateful to Dr. Sally Solomon, Dr. Caroline Schauer and Dr. Jian-Min Yuan for their detailed comments on my oral proposal and thesis. I would like to thank many of my collaborators. I specially thank Dr. Jian-Min Yuan for his valuable suggestion and assistance, especially in protein folding and amyloid β aggregation projects. I also would like to thank Dr. Karl Sohlberg for giving me the opportunities to work with him on an important project. I would also thank Dr. Reinhard Schweitzer-Stenner for his wonderful insights on many of my research works, especially on the project of aggregation of Amyloid β peptide. I also thank Dr. Solomon Praveen for 4 his help in dental materials projects and suggestions in thesis writing. Many thanks are due to Dr. Thomas G. Spiro of Princeton University and his student Dr. Gurusamy Balakrishnan on assistance in Raman spectroscopy. I am grateful to many people for their selfless support in many areas. I especially acknowledge Dr. Patrick Loll for his assistance in circular dichroism spectroscopy, Dr. Guoliang Yang for AFM studies, Ms. Edith Smith for her kindly help and coordination in obtaining the chemicals and instruments for our research work. Many thanks are due to my friends in the Department of Chemistry who make my life here memorable. I thank Dr. Shuxi Li, Dr. Qiuwei Feng, Dr. Shan Cheng, Dr. Hua Dong, Dr. Houping Yin, Ms. Alpa Patel, Mr. Yi Guo, Dr. Jim Tu, Ms. Stephanie Schuster for their collaboration, discussions and help during these years. I also want to thank all the professors, staff and students in Department of Chemistry, Drexel University for making the Department such a joyful working and studying environment. Finally, I am greatly grateful to my parents, Mr. Chongzhen Sun and Mrs. Xiuqing Nie, for their continuous encouragement and unconditional love throughout in my life. 5 Table of Contents Table of Contents................................................................................................................ 6 List of Tables .................................................................................................................... 15 List of Figures................................................................................................................... 17 Abstract............................................................................................................................. 24 Chapter 1: An Overview to Nanoporous Sol-Gel Materials............................................. 27 1.1 Introduction.................................................................................................. 27 1.2 Fundamentals of Sol-Gel Process ................................................................ 29 1.2.1 Sol-Gel Reactions .................................................................................... 30 1.3 Nanoporous Sol-Gel Materials from Surfactant Templated Pathways........ 31 1.4 Nanoporous Sol-Gel Materials from Nonsurfactant Templated Pathway ... 35 1.5 Characterizations of Nanoporous Materials................................................. 37 1.5.1 Gas sorption measurement.................................................................... 37 1.5.2 X-Ray Diffraction (XRD)..................................................................... 40 1.5.3 Electron Microscopy............................................................................. 41 1.6 Organic/Inorganic Hybrid Materials by Sol-Gel Approach......................... 42 1.7 Sol-Gel Encapsulation of Biomolecules ...................................................... 45 1.8 References.................................................................................................... 49 Chapter 2: Rigid Matrix Artificial Chaperon (RMAC) – Mediated Refolding of Cytochrome c.................................................................................................................... 63 2.1 Introduction......................................................................................................... 63 2.1.1 Fundamentals of Protein Folding Unfolding ........................................... 63 2.1.2 Chaperone and Protein Folding ............................................................... 64 6 2.1.3 Rigid Matrix Artificial Chaperone (RMAC) ........................................... 65 2.1.4 Why Cytochrome c? ................................................................................ 67 2.1.5 Analytical Methods to Monitor Cytochrome c’s Folding Unfolding ...... 68 2.1.5.1 Fluorescence Spectroscopy........................................................... 68 2.1.5.2 Circular Dichroism (CD) .............................................................. 69 2.2 Experimental....................................................................................................... 71 2.2.1 Materials .................................................................................................. 71 2.2.2 Encapsulation of Unfolded Cc into Silica Matrix.................................... 71 2.2.3 Removal of Templates ............................................................................. 73 2.2.4 Characterizations of Nanoporous Silica Matrix....................................... 73 2.2.5 Fluorescence Spectroscopy of Encapsulated Cc...................................... 73 2.2.6 Circular Dichroism of Cc......................................................................... 74 2.2.7 Fourier Transform Infrared Spectroscopy (FTIR) of Silica Matrix......... 75 2.2.8 UV-Vis Spectroscopy (UV) of Encapsulated Cc..................................... 76 2.2.9 Attempts of Making Silica Thin Film...................................................... 77 2.3 Results and Discussion ....................................................................................... 78 2.3.1 Characterization of Silica Matrix............................................................. 79 2.3.2 FT-IR Spectroscopy on Silica Matrix...................................................... 80 2.3.3 Fluorescence Spectroscopy of Encapsulated Cc...................................... 80 2.3.4 Leakage Tests........................................................................................... 81 2.3.5 CD Spectroscopy ..................................................................................... 82 2.4 Conclusion .......................................................................................................... 83 2.5 Acknowledgement .............................................................................................. 83 7 2.6 References........................................................................................................... 83 Chapter 3: Using Resonance Raman Spectroscopy to Study Folding Unfolding Behavior of Encapsulated Heme Proteins in Silica Matrix with Controlled Pore Sizes .................. 99 3.1 Introduction......................................................................................................... 99 3.1.1 Resonance Raman Spectroscopy and Protein Folding............................. 99 3.1.2 Heme Protein ......................................................................................... 102 3.1.2.1 Cytochrome c and its Resonance Raman Spectroscopy ............. 102 3.1.2.1.1 Marker Band Region........................................................ 103 3.1.2.1.2 Fingerprint Region........................................................... 104 3.1.2.2 Hemoglobin (Hb) and Its Resonance Raman Spectra................. 104 3.1.2.3 Myoglobin (Mb) and Its Resonance Raman Spectra .................. 105 3.1.2.4 Protein Encapsulation ................................................................. 108 3.2 Experimental..................................................................................................... 109 3.2.1 Materials ...............................................................................................
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