Xu, Guolin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 111-128). / Abstracts in English and Chinese. / ABSTRACT --- p.I / ACKNOWLEDGEMENTS --- p.IV / LIST OF FIGURES --- p.VII / ABBBREVIATIONS --- p.X / Chapter CHAPTER ONE --- INTRODUCTION --- p.1 / Chapter 1.1 --- Antigens --- p.1 / Chapter 1.1.1 --- Preamble --- p.1 / Chapter 1.1.2 --- Types of antigen --- p.1 / Chapter 1.1.3 --- Autoantigens --- p.2 / Chapter 1.1.4 --- Telomerase is an important autoantigen --- p.6 / Chapter 1.2 --- Antibodies --- p.12 / Chapter 1.2.1 --- Preamble --- p.12 / Chapter 1.2.2 --- Ig structure --- p.12 / Chapter 1.2.3 --- Ig synthesis --- p.13 / Chapter 1.2.4 --- Immunoglobulin isotypes --- p.15 / Chapter 1.2.5 --- Monoclonal antibodies (mAb) --- p.17 / Chapter 1.2.6 --- Autoantibodies --- p.20 / Chapter 1.2.7 --- Telomerase detection and antibodies to telomerase --- p.22 / Chapter 1.3 --- Object and Scope of Study --- p.24 / Chapter CHAPTER TWO --- MATERIALS AND METHODS --- p.31 / Chapter 2.1 --- Materials --- p.31 / Chapter 2.1.1 --- Animals --- p.31 / Chapter 2.1.2 --- Antibodies --- p.31 / Chapter 2.1.3 --- Primers --- p.31 / Chapter 2.1.4 --- Culture media and reagents --- p.32 / Chapter 2.1.5 --- Chemicals and enzymes --- p.32 / Chapter 2.1.6 --- Miscellaneous chemicals --- p.33 / Chapter 2.1.7 --- Commercial kits --- p.33 / Chapter 2.1.8 --- Instruments --- p.33 / Chapter 2.1.9 --- Buffers --- p.34 / Chapter 2.2 --- Methods --- p.35 / Chapter 2.2.1 --- Cells and cell culture --- p.35 / Chapter 2.2.2 --- Polymerase chain reaction (PCR) --- p.36 / Chapter 2.2.3 --- Cloning of C-terminal gene fragment to pGEX cloning vector --- p.36 / Chapter 2.2.4 --- Detection of antibody activity by ELISA --- p.37 / Chapter 2.2.5 --- Histochemical Staining --- p.38 / Chapter 2.2.6 --- Hybridoma production --- p.40 / Chapter 2.2.7 --- Protein analysis --- p.43 / Chapter 2.2.8 --- Flow cytometry --- p.46 / Chapter 2.2.9 --- Animal handling --- p.47 / Chapter 2.2.10 --- Statistical analysis --- p.48 / Chapter CHAPTER THREE --- PRELIMINARY STUDIES USING THE ANTI-N-TERT-TELOMERASE MAB DERIVED FROM HYBRIDOMA 476 --- p.49 / Chapter 3.1 --- Preamble --- p.49 / Chapter 3.2 --- Hybridoma 476 cells can be stained by the labeled recombinant N-TERT antigen --- p.50 / Chapter 3.3 --- Mouse spleen cells can also be stained by biotin-labeled N-TERT antigen --- p.52 / Chapter 3.4 --- Discussion --- p.53 / Chapter CHAPTER FOUR --- PRODUCTION OF MONOCLONAL ANTIBODIES TO C-TERT --- p.63 / Chapter 4.1 --- Preamble --- p.63 / Chapter 4.2 --- Construction of C-TERT expression vector --- p.63 / Chapter 4.3 --- Expression and purification of recombinant human C-terminal telomerase antigen --- p.64 / Chapter 4.4 --- Immunization of Balb/c mice with C-TERT-GST --- p.65 / Chapter 4.5 --- Generation of hybridomas to C-TERT --- p.65 / Chapter 4.6 --- Identification and selection of reactive clones --- p.65 / Chapter CHAPTER FIVE --- CHARACTERIZATION OF MAB A63 --- p.71 / Chapter 5.1 --- Preamble --- p.71 / Chapter 5.2 --- Characterization of mAb A63 by ELISA --- p.71 / Chapter 5.3 --- Characterization of mAb A63 by Western blotting analysis --- p.73 / Chapter 5.4 --- Characterization of mAb A63 by immuno-histochemical staining --- p.73 / Chapter 5.5 --- mAb A63 can also stain fish telomerase and human placenta --- p.75 / Chapter 5.6 --- mAb A63 can also stain telomerase in human tumors --- p.76 / Chapter 5.7 --- Hybridoma A63 can produce ascites fluid --- p.76 / Chapter 5.8 --- Discussion --- p.77 / Chapter CHAPTER SIX --- GENERAL DISCUSSION --- p.93 / Chapter 6.1 --- Why Enhancing buffer is required for the nuclear staining of hTERT when using mAb 476 or mAb A63 --- p.96 / Chapter 6.2 --- Why hybridoma 476 failed to form ascites while hybridoma A63 succeeded --- p.99 / Chapter 6.3 --- Can IL-6 be used to treat autoimmune diseases? --- p.102 / Chapter 6.4 --- Possible use of monoclonal antibodies in cancer therapy --- p.104 / Chapter 6.5 --- Prospects on study --- p.107 / REFERENCES --- p.111
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326656 |
| Date | January 2009 |
| Contributors | Xu, Guolin., Chinese University of Hong Kong Graduate School. Division of Chemical Pathology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xi, 128 leaves : ill. ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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