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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77281完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張世宗 | zh_TW |
| dc.contributor.author | 許翔輔 | zh_TW |
| dc.contributor.author | Hsiang-Fu Hsu | en |
| dc.date.accessioned | 2021-07-10T21:54:02Z | - |
| dc.date.available | 2024-08-13 | - |
| dc.date.copyright | 2019-08-14 | - |
| dc.date.issued | 2019 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | 1. BR, D. G. (1994) Structures of Botulinum neurotoxin, its functional domains and perspectives on the crystalline type A toxin. Therapy with Botulinum Toxin, 15-39
2. Snipe, P. T., and Sommer, H. (1928) Studies on Botulinus Toxin: 3. Acid Precipitation of Botulinus Toxin [with Discussion]. The Journal of Infectious Diseases 43, 152-160 3. Edward J. Schantz, E. A. J. (1997) Botulinum Toxin: The Story of Its Development for the Treatment of Human Disease. Perspectives in Biology and Medicine 40, 11 4. Nigam, P. K., and Nigam, A. (2010) Botulinum toxin. Indian journal of dermatology 55, 8-14 5. M.D. Collins, A. K. E. (1988) Phylogeny and taxonomy of the food-borne pathogen Clostridium botulinum and its neurotoxins. Journal of Applied Microbiology 84, 5-17 6. Hill, K. K., Smith, T. J., Helma, C. H., Ticknor, L. O., Foley, B. T., Svensson, R. T., Brown, J. L., Johnson, E. A., Smith, L. A., Okinaka, R. T., Jackson, P. J., and Marks, J. D. (2007) Genetic diversity among Botulinum Neurotoxin-producing clostridial strains. J Bacteriol 189, 818-832 7. Pellett, S., Yaksh, L. T., and Ramachandran, R. (2015) Current Status and Future Directions of Botulinum Neurotoxins for Targeting Pain Processing. Toxins 7 8. Lacy, D. B., and Stevens, R. C. (1999) Sequence homology and structural analysis of the clostridial neurotoxins 11 Edited by G. Von Heijne. Journal of Molecular Biology 291, 1091-1104 9. Montal, M. (2010) Botulinum neurotoxin: a marvel of protein design. Annu Rev Biochem 79, 591-617 10. Schiavo, G., Matteoli, M., and Montecucco, C. (2000) Neurotoxins Affecting Neuroexocytosis. Physiological Reviews 80, 717-766 11. Schiavo, G., Poulain, B., Rossetto, O., Benfenati, F., Tauc, L., and Montecucco, C. (1992) Tetanus toxin is a zinc protein and its inhibition of neurotransmitter release and protease activity depend on zinc. EMBO J 11, 3577-3583 12. Inoue, K., Fujinaga, Y., Watanabe, T., Ohyama, T., Takeshi, K., Moriishi, K., Nakajima, H., Inoue, K., and Oguma, K. (1996) Molecular composition of Clostridium botulinum type A progenitor toxins. Infection and Immunity 64, 1589-1594 13. Fujinaga, Y., Inoue, K., Watarai, S., Sakaguchi, Y., Arimitsu, H., Lee, J.-C., Jin, Y., Matsumura, T., Kabumoto, Y., Watanabe, T., Ohyama, T., Nishikawa, A., and Oguma, K. (2004) Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes. Microbiology 150, 1529-1538 14. Scott, A. B. (1980) Botulinum Toxin Injection into Extraocular Muscles as an Alternative to Strabismus Surgery. Ophthalmology 87, 1044-1049 15. Carruthers, J. D. A., and Carruthers, J. A. (1992) Treatment of Glabellar Frown Lines with C. Botulinum-A Exotoxin. The Journal of Dermatologic Surgery and Oncology 18, 17-21 16. Caleo, M., and Schiavo, G. (2009) Central effects of tetanus and botulinum neurotoxins. Toxicon 54, 593-599 17. Foran, P. G., Mohammed, N., Lisk, G. O., Nagwaney, S., Lawrence, G. W., Johnson, E., Smith, L., Aoki, K. R., and Dolly, J. O. (2003) Evaluation of the Therapeutic Usefulness of Botulinum Neurotoxin B, C1, E, and F Compared with the Long Lasting Type A: BASIS FOR DISTINCT DURATIONS OF INHIBITION OF EXOCYTOSIS IN CENTRAL NEURONS. Journal of Biological Chemistry 278, 1363-1371 18. Shoemaker, C. B., and Oyler, G. A. (2013) Persistence of Botulinum Neurotoxin Inactivation of Nerve Function. in Botulinum Neurotoxins (Rummel, A., and Binz, T. eds.), Springer Berlin Heidelberg, Berlin, Heidelberg. pp 179-196 19. Frevert, J. (2010) Content of botulinum neurotoxin in Botox®/Vistabel®, Dysport®/Azzalure®, and Xeomin®/Bocouture®. Drugs R D 10, 67-73 20. Torres, S., Hamilton, M., Sanches, E., Starovatova, P., Gubanova, E., and Reshetnikova, T. (2013) Neutralizing antibodies to botulinum neurotoxin type A in aesthetic medicine: five case reports. Clinical, cosmetic and investigational dermatology 7, 11-17 21. Dressler, D. (2008) Botulinum toxin drugs: future developments. Journal of Neural Transmission 115, 575-577 22. Kukreja, R., Chang, T.-W., Cai, S., Lindo, P., Riding, S., Zhou, Y., Ravichandran, E., and Singh, B. R. (2009) Immunological characterization of the subunits of type A botulinum neurotoxin and different components of its associated proteins. Toxicon 53, 616-624 23. Fabbri, M., Leodori, G., Fernandes, R. M., Bhidayasiri, R., Marti, M. J., Colosimo, C., and Ferreira, J. J. (2016) Neutralizing Antibody and Botulinum Toxin Therapy: A Systematic Review and Meta-analysis. Neurotoxicity Research 29, 105-117 24. Hefter, H., Hartmann, C., Kahlen, U., Moll, M., and Bigalke, H. (2012) Prospective analysis of neutralising antibody titres in secondary non-responders under continuous treatment with a botulinumtoxin type A preparation free of complexing proteins—a single cohort 4-year follow-up study. BMJ Open 2, e000646 25. Göschel, H., Wohlfarth, K., Frevert, J., Dengler, R., and Bigalke, H. (1997) Botulinum A Toxin Therapy: Neutralizing and Nonneutralizing Antibodies—Therapeutic Consequences. Experimental Neurology 147, 96-102 26. Birklein, F., and Erbguth, F. (2000) Sudomotor testing discriminates between subjects with and without antibodies against botulinum toxin A—A preliminary observation. Movement Disorders 15, 146-149 27. Voller, B., Moraru, E., Auff, E., Benesch, M., Poewe, W., Wissel, J., Müller, J., Entner, T., Bigalke, H., and Schnider, P. (2004) Ninhydrin sweat test: A simple method for detecting antibodies neutralizing botulinum toxin type A. Movement Disorders 19, 943-947 28. Hanna, P. A., Jankovic, J., and Vincent, A. (1999) Comparison of mouse bioassay and immunoprecipitation assay for botulinum toxin antibodies. Journal of Neurology, Neurosurgery & Psychiatry 66, 612-616 29. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248-254 30. Costa, S., Almeida, A., Castro, A., and Domingues, L. (2014) Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system. Front Microbiol 5, 63-63 31. LaVallie, E. R., DiBlasio, E. A., Kovacic, S., Grant, K. L., Schendel, P. F., and McCoy, J. M. (1993) A Thioredoxin Gene Fusion Expression System That Circumvents Inclusion Body Formation in the E. coli Cytoplasm. Bio/Technology 11, 187-193 32. Mousavi, M. L., Montaser Kouhsari, S., Nazarian, S., Rasooli, I., and Amani, J. (2004) Cloning, expression and purification of Clostridium botulinum neurotoxin type E binding domain. Iranian Journal of Biotechnology 2, 183-188 33. Davis, G. D., Elisee, C., Newham, D. M., and Harrison, R. G. (1999) New fusion protein systems designed to give soluble expression in Escherichia coli. Biotechnology and Bioengineering 65, 382-388 34. Baldwin, M. R., Bradshaw, M., Johnson, E. A., and Barbieri, J. T. (2004) The C-terminus of botulinum neurotoxin type A light chain contributes to solubility, catalysis, and stability. Protein Expression and Purification 37, 187-195 | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77281 | - |
| dc.description.abstract | 肉毒桿菌 (Clostridium botulinum) 可生產八種具不同特性的外毒素 (type A, B, C1, C2, D, E, F, and G)，並作用於神經肌肉會合處阻礙乙醯膽鹼釋放，以中斷神經訊號傳導，造成肌肉癱瘓。雖然肉毒桿菌素 (botulinum neurotoxin) 是自然界中毒性最強的物質之一，但是調整使用量後，則可以應用於治療斜視、面肌痙攣與某些類型的語言障礙等。此外，由於其低副作用與高耐受性，更使得注射肉毒桿菌素成為流行的醫美項目，然而長時間接受肉毒桿菌素注射，可能會因患者產生中和性抗體而降低效果甚至失效，因此有必要發展一種評估患者對肉毒桿菌素抗性的工具。現行用於醫療與醫美主要是A型肉毒桿菌素，本研究於大腸桿菌中分別表現A型肉毒桿菌素的受體結合區 (receptor-binding domain)、易位區 (translocation domain) 與肽酶區 (peptidase domain)，再以管柱層析法進行純化，以建立可以偵測抗A型肉毒桿菌素抗體之酵素聯結免疫吸附分析法。本研究成果有助於檢測患者之抗藥性生成情形，並藉此調整治療方法及避免無效醫療。 | zh_TW |
| dc.description.abstract | Clostridium botulinum produces eight antigenically distinctive exotoxins (type A, B, C1, C2, D, E, F, and G), which can block the release of acetylcholine and the neurotransmission at the neuromuscular junction for causing muscle paralysis. Though botulinum neurotoxin (BTX) is one of the most poisonous biological substances known, it has also been used as therapies for strabismus, hemifacial spasm, some types of language disorder, and the cosmetic surgery after dose adjustment. The few side effects and well tolerability made injection of neurotoxin a popular cosmetological application. However, in long-term BTX treatment, loss of therapeutic efficacy may occur due to induction of the neutralizing antibody, which causes non-responsiveness to treatment. Therefore, it’s necessary to develop a method for dermatologists to evaluate the efficacy of BTX before injection in patients.
The BTX type A has been widely used for medical and cosmetological application. In this study, the receptor-binding domain, translocation domain and peptidase domain of BTX type A were expressed in E. coli, and purified by column chromatography. The protein domains of BTX were applied in establishing an enzyme-linked immunosorbent assay for detecting the anti-neurotoxin antibody. The present study developed a useful tool for monitoring the formation of the anti-neurotoxin antibody in patients and can be considered as a diagnostic index for preventing futile medical cares. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T21:54:02Z (GMT). No. of bitstreams: 1 ntu-108-R06b22043-1.pdf: 1771966 bytes, checksum: 82a738427676d1d1df0763bfc82f4c96 (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 目錄 i
摘要 iv Abstract v 縮寫表 vi 第一章 緒論 1 1.1 肉毒桿菌的簡介 1 1.1.1肉毒桿菌的發現 1 1.1.2 肉毒桿菌的表現型分類 1 1.1.3 肉毒桿菌中毒 2 1.2 肉毒桿菌素簡介 2 1.2.1 肉毒桿菌素的分類 2 1.2.2 肉毒桿菌素的結構與作用機制 3 1.2.3 肉毒桿菌素關聯蛋白 3 1.2.4 肉毒桿菌素的醫學用途 4 1.2.5 肉毒桿菌素的效期 4 1.2.6 肉毒桿菌素商品與基因來源 5 1.3 抗肉毒桿菌素之抗體 5 1.3.1 肉毒桿菌素的中和性抗體 5 1.3.2 肉毒桿菌素的免疫原性 5 1.3.3 肉毒桿菌素抗體效價測試 6 1.4 酵素聯結免疫吸附分析法之類型 7 1.4.1 直接型ELISA (Direct ELISA) 7 1.4.2 非直接型ELISA (Indirect ELISA) 7 1.4.3 三明治型ELISA (Sandwich ELISA) 7 1.4.4 競爭型ELISA (Competitive ELISA) 8 1.5 研究動機與目的 8 第二章 材料與方法 9 2.1 實驗材料 9 2.1.1 大腸桿菌菌株 9 2.2 BTX表現載體之建構 9 2.2.1 大腸桿菌表現載體 9 2.2.2 核酸引子設計 10 2.2.3 聚合酶連鎖反應 10 2.2.4 限制酶切反應 10 2.2.5 接合反應 11 2.2.6 轉形作用 11 2.3 DNA實驗方法 11 2.3.1 質體DNA製備 11 2.3.2 洋菜膠電泳 12 2.3.3 DNA片段之純化 12 2.3.4 DNA定量 12 2.4 蛋白質實驗方法 12 2.4.1 蛋白質表現 12 2.4.2 Ni-NTA agarose親和層析法 13 2.4.3離子交換法 13 2.4.4 蛋白質定量 13 2.4.5 蛋白質電泳 14 2.4.6 蛋白質電泳膠片染色 14 2.4.7 蛋白質電泳轉印 14 2.4.8 西方墨點法 15 2.4.9 蛋白質脫鹽與濃縮 15 第三章 結果 16 3.1 肉毒桿菌素蛋白質區塊BTX-RB表現載體之建構 16 3.2 肉毒桿菌素蛋白質區塊BTX-RB於大腸桿菌之表現 16 3.3 BTX-RB重組蛋白質之純化 16 3.4 肉毒桿菌素蛋白質區塊BTX-P與BTX-T表現載體之建構 17 3.5 肉毒桿菌素蛋白質區塊BTX-P之純化 18 3.6 以連續性梯度純化肉毒桿菌素蛋白質區塊BTX-T 18 3.7 以非連續性梯度純化肉毒桿菌素蛋白質區塊BTX-T 19 第四章 討論 20 4.1 肉毒桿菌素受體連接區BTX-RB之表現與純化方法探討 20 4.2 肉毒桿菌素肽酶區BTX-P之表現及純化方法探討 20 4.3 肉毒桿菌素易位區BTX-T之表現與純化方法探討 21 4.4 肉毒桿菌素抗體偵測平台建立之探討 21 參考文獻 22 表與圖 26 圖一 A型肉毒桿菌素蛋白質結構圖 27 圖二 BTX-RB重組大腸桿菌表現載體之建構 28 圖三 使用不同載體以大腸桿菌系統表現BTX-RB 29 圖四 以pET28a載體表現BTX-RB重組蛋白 30 圖五 利用質譜儀分析，鑑定重組BTX-RB之身分 31 圖六 以pET30a載體表現BTX-RB 32 圖七 BTX-T與BTX-P重組大腸桿菌表現載體之建構 33 圖八 以pET30a載體表現BTX-P 34 圖九 以HitrapTM DEAE FF管柱純化BTX-P 35 圖十 以pET30a載體表現BTX-T 36 圖十一 以非連續性梯度純化BTX-T 37 圖十二 以HitrapTM DEAE FF管柱純化BTX-T 38 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 肉毒桿菌素 | zh_TW |
| dc.subject | 酵素聯結免疫吸附分析法 | zh_TW |
| dc.subject | botulinum neurotoxin | en |
| dc.subject | enzyme-linked immunosorbent assay | en |
| dc.title | 表現肉毒桿菌素之蛋白質區塊以應用於臨床檢測抗肉毒桿菌素之抗體 | zh_TW |
| dc.title | Expression of the Protein Domains of Clostridium botulinum Neurotoxin for Applying in Clinical Detection of Anti-Neurotoxin Antibody | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 107-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 陳威戎;林翰佳;黃純芳 | zh_TW |
| dc.contributor.oralexamcommittee | ;; | en |
| dc.subject.keyword | 肉毒桿菌素,酵素聯結免疫吸附分析法, | zh_TW |
| dc.subject.keyword | botulinum neurotoxin,enzyme-linked immunosorbent assay, | en |
| dc.relation.page | 38 | - |
| dc.identifier.doi | 10.6342/NTU201902730 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2019-08-12 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生化科技學系 | - |
| 顯示於系所單位： | 生化科技學系 | |
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