請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81284完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳林祈(Lin-Chi Chen) | |
| dc.contributor.author | You-Cheng Lin | en |
| dc.contributor.author | 林佑承 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:40:45Z | - |
| dc.date.available | 2021-09-01 | |
| dc.date.available | 2022-11-24T03:40:45Z | - |
| dc.date.copyright | 2021-09-01 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-08-25 | |
| dc.identifier.citation | 黃惟甄。2020。黏蛋白-1適體篩選與分析。碩士論文。台北：國立台灣大學生物機電工程學系研究所。 Abdulhalim, I., Zourob, M., Lakhtakia, A. (2008). Surface Plasmon Resonance for Biosensing: A Mini-Review. Electromagnetics, 28(3), 214-242 Ali, M. H., Elsherbiny, M. E., Emara, M. (2019). Updates on Aptamer Research. Int J Mol Sci, 20(10), 2511 Baouendi, M., Cognet, J. A., Ferreira, C. S., Missailidis, S., Coutant, J., Piotto, M., Hantz, E., Hervé du Penhoat, C. (2012). Solution structure of a truncated anti‐MUC1 DNA aptamer determined by mesoscale modeling and NMR. The FEBS journal, 279(3), 479-490. Da Pieve, C., Williams, P., Haddleton, D. M., Palmer, R. M., Missailidis, S. (2010). Modification of thiol functionalized aptamers by conjugation of synthetic polymers. Bioconjugate chemistry, 21(1), 169-174. Ferreira, C. S., Cheung, M. C., Missailidis, S., Bisland, S., Gariépy, J. (2009). Phototoxic aptamers selectively enter and kill epithelial cancer cells. Nucleic acids research, 37(3), 866-876. Ferreira, C. S. M., Matthews, C. S., Missailidis, S. (2006). DNA aptamers that bind to MUC1 tumour marker: design and characterization of MUC1-binding single-stranded DNA aptamers. Tumor biology, 27(6), 289-301 Ferreira, C. S. M., Papamichael, K., Guilbault, G., Schwarzacher, T., Gariepy, J., Missailidis, S. (2008). DNA aptamers against the MUC1 tumour marker: design of aptamer–antibody sandwich ELISA for the early diagnosis of epithelial tumours. Analytical and bioanalytical chemistry, 390(4), 1039-1050. Guillaume, T., Dehame, V., Chevallier, P., Peterlin, P., Garnier, A., Grégoire, M., Pichinuk, E., Rubinstein, D. B., Wreschner, D. H. (2019). Targeting cell-bound MUC1 on myelomonocytic, monocytic leukemias and phenotypically defined leukemic stem cells with anti-SEA module antibodies. Experimental hematology, 70, 97-108. Gupta, P., Bharti, A., Kaur, N., Singh, S., Prabhakar, N. (2018). An electrochemical aptasensor based on gold nanoparticles and graphene oxide doped poly (3, 4-ethylenedioxythiophene) nanocomposite for detection of MUC1. Journal of Electroanalytical Chemistry, 813, 102-108. Hianik, T., Ostatná, V., Sonlajtnerova, M., Grman, I. (2007). Influence of ionic strength, pH and aptamer configuration for binding affinity to thrombin. Bioelectrochemistry, 70(1), 127-133. Huang, Z., Szostak, J. W. (2003). Evolution of aptamers with a new specificity and new secondary structures from an ATP aptamer. Rna, 9(12), 1456-1463. Le, T. T., Chumphukam, O., Cass, A. E. (2014). Determination of minimal sequence for binding of an aptamer. A comparison of truncation and hybridization inhibition methods. RSC Advances, 4(88), 47227-47233. Levitin, F., Stern, O., Weiss, M., Gil-Henn, C., Ziv, R., Prokocimer, Z., Smorodinsky, N. I., Rubinstein, D. B., Wreschner, D. H. (2005). The MUC1 SEA module is a self-cleaving domain. Journal of Biological Chemistry, 280(39), 33374-33386. Liu, X., Qin, Y., Deng, C., Xiang, J., Li, Y. (2015). A simple and sensitive impedimetric aptasensor for the detection of tumor markers based on gold nanoparticles signal amplification. Talanta, 132, 150-154. Macao, B., Johansson, D. G., Hansson, G. C., Härd, T. (2006). Autoproteolysis coupled to protein folding in the SEA domain of the membrane-bound MUC1 mucin. Nature structural molecular biology, 13(1), 71-76. Mayer, G. (2009). The chemical biology of aptamers. Angewandte Chemie International Edition, 48(15), 2672-2689. Meirinho, S. G., Dias, L. G., Peres, A. M., Rodrigues, L. R. (2016). Voltammetric aptasensors for protein disease biomarkers detection: A review. Biotechnology advances, 34(5), 941-953. Mehlhorn, A., Rahimi, P., Joseph, Y. (2018). Aptamer-based biosensors for antibiotic detection: a review. Biosensors, 8(2), 54. Nabavinia, M. S., Gholoobi, A., Charbgoo, F., Nabavinia, M., Ramezani, M., Abnous, K. (2017). Anti‐MUC1 aptamer: a potential opportunity for cancer treatment. Medicinal Research Reviews, 37(6), 1518-1539. Nawaz, M. A. H., Rauf, S., Catanante, G., Nawaz, M. H., Nunes, G., Marty, J. L., Hayat, A. (2016). One step assembly of thin films of carbon nanotubes on screen printed interface for electrochemical aptasensing of breast cancer biomarker. Sensors, 16(10), 1651. Paimard, G., Shahlaei, M., Moradipour, P., Karamali, V., Arkan, E. (2020). Impedimetric aptamer based determination of the tumor marker MUC1 by using electrospun core-shell nanofibers. Microchimica Acta, 187(1), 1-10. Paniel, N., Baudart, J., Hayat, A., Barthelmebs, L. (2013). Aptasensor and genosensor methods for detection of microbes in real world samples. Methods, 64(3), 229-240. Perret, G., Boschetti, E. (2019). Aptamer-based affinity chromatography for protein extraction and purification. Aptamers in Biotechnology, 93-139. Pichinuk, E., Benhar, I., Jacobi, O., Chalik, M., Weiss, L., Ziv, R., Sympson, C., Karwa, A., Smorodinsky, N. I., Rubinstein, D. B., Wreschner, D. H. (2012). Antibody targeting of cell-bound MUC1 SEA domain kills tumor cells. Cancer research, 72(13), 3324-3336. Prieto-Simón, B., Campàs, M., Marty, J. L. (2010). Electrochemical aptamer-based sensors. Bioanalytical Reviews, 1(2-4), 141-157. Radi, A. E. (2011). Electrochemical aptamer-based biosensors: recent advances and perspectives. International journal of electrochemistry, 2011. Singh, R., Bandyopadhyay, D. (2007). MUC1: A target molecule for cancer therapy.Cancer biology therapy, 6, 481-486 Slavkovic, S., Eisen, S. R., Johnson, P. E. (2020). Designed alteration of binding affinity in structure-switching aptamers through the use of dangling nucleotides. Biochemistry, 59(5), 663-670. Vu, C. Q., Rotkrua, P., Tantirungrotechai, Y., Soontornworajit, B. (2017). Oligonucleotide hybridization combined with competitive antibody binding for the truncation of a high-affinity aptamer. ACS combinatorial science, 19(10), 609-617. Walter, J. G., Stahl, F., Scheper, T. (2012). Aptamers as affinity ligands for downstream processing. Engineering in Life Sciences, 12(5), 496-506. Yang, D. K., Chou, C. F., Chen, L. C. (2018). Selection of aptamers for AMACR detection from DNA libraries with different primers. RSC advances, 8(34), 19067-19074. Yousefi, M., Dehghani, S., Nosrati, R., Zare, H., Evazalipour, M., Mosafer, J., Ramezani, M. (2019). Aptasensors as a new sensing technology developed for the detection of MUC1 mucin: A review. Biosensors and Bioelectronics, 130, 1-19. Zhao, Q., Wu, M., Le, X. C., Li, X. F. (2012). Applications of aptamer affinity chromatography. TrAC Trends in Analytical Chemistry, 41, 46-57. Zon, G. (2020). 'Recent advances in aptamer applications for analytical biochemistry.' Analytical Biochemistry: 113894. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81284 | - |
| dc.description.abstract | "黏蛋白-1(mucin-1；MUC1)是人類癌症研究的重要生物標記，在常見癌症，如肺腺癌、乳癌等都會有過度表現的狀況。在臨床上以MUC1 SEA單元作為標靶治療，若能快速且精準的偵測此單元，將能對癌症的早期診斷有偌大的益處。近年來發展適體感測器用於即時檢測MUC1，其具備高靈敏度及與MUC1高結合力等優勢。為了達成高效感測的目標，除了提升適體的效能、優化感測條件外，並且可以使用抗體輔助來提升適體感測器之性能。針對實驗室已篩選出MUC1 SEA單元(從未有篩選過)的K2適體進行優化與感測應用。首先，透過表面電漿共振分析證明K2適體與MUC1 SEA單元有專一性的結合，接著使用熔解曲線分析對K2適體的結構進行穩定性分析，透過溫度和離子強度的調整，找到最適的結合條件為2 mM MgCl2 及100 mM NaCl應用於電化學適體感測器分析，而且發現K2適體和抗體各別與MUC1 SEA單元有不同的結合位點。整合K2適體的最適化條件且與抗體結合MUC1 SEA單元的位點不同，將其應用於抗體輔助電化學適體感測器檢測MUC1 SEA單元，使用電化學阻抗頻譜進行感測。實驗結果顯示在只有K2適體的電化學適體感測器中，檢測MUC1 SEA單元的濃度線性區間在29.6至66.6 nM之間且檢測極限在8.59 nM，此感測器計算得到的KD,app值為77.6 nM，R2為0.904。另外，透過使用抗體輔助電化學感測器測量到MUC1 SEA單元的濃度線性區間為1.23至11.1 nM且檢測極限為0.48 nM，此感測器計算得到的KD,app值為33.8 nM，R2為0.977，綜合兩種感測器的數據結果比較，以抗體輔助電化學適體感測器顯著地提升檢測濃度範圍及偵測極限，也證明其對MUC1 SEA 單元作為標靶檢測具有潛力。" | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:40:45Z (GMT). No. of bitstreams: 1 U0001-2307202119082000.pdf: 4894370 bytes, checksum: f719f3ae3a41653114200b744bdfc4c8 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 致謝 i 摘要 ii Abstract iii 目錄 v 圖目錄 viii 表目錄 xii 符號說明 xiii 第一章 緒論 1 1.1 研究背景 1 1.2 研究動機 3 1.3 研究目的 4 1.4 研究架構 5 第二章 文獻探討 6 2.1 MUC1 SEA單元 6 2.2 適體的結構穩定性與結合力 8 2.2.1 鹼基差異對適體結構的影響 8 2.2.2 環境因子對適體結合力的影響 12 2.3表面電漿共振分析 14 2.3.1實驗原理與反應機制 14 2.3.2 MUC1適體的動力學參數 21 2.4 MUC1適體感測回顧 24 第三章 研究方法 30 3.1 實驗儀器與藥品 30 3.1.1 實驗儀器 30 3.1.2 實驗藥品 31 3.2 實驗操作方法及步驟 32 3.2.1 酶聯寡核苷酸分析法 32 3.2.2 表面電漿共振分析 35 3.2.3 熔解曲線分析 38 3.2.4 電化學適體感測器 40 第四章 實驗結果與討論 43 4.1 適體序列優化 43 4.1.1 K2適體的核心區域鹼基亂數對MUC1 SEA單元結合力的影響 43 4.1.2 K2適體的引子截斷對MUC1 SEA單元結合力的影響 47 4.1.3 K2適體的動力學分析 51 4.1.4 K2-de5’適體的動力學分析 54 4.1.5 K2適體和K2-de5’適體的專一性比較 56 4.1.6 小結 58 4.2 適體環境優化 59 4.2.1 溫度對K2適體的結合力影響 59 4.2.2 溶液離子濃度對K2適體的結合力影響 62 4.2.3 溶液離子濃度對K2適體的結構影響 64 4.2.4 小結 66 4.3 適體與抗體競爭結合分析 67 4.3.1 SPR分析測定結合位點 67 4.3.2微盤分析儀測定結合位點 69 4.3.3 小結 71 4.4 電化學阻抗式適體感測MUC1 SEA單元 72 4.4.1靈敏度感測分析 72 4.4.2專一性感測分析 81 4.4.3 小結 87 4.5 抗體輔助電化學阻抗式適體感測MUC1 SEA單元 88 4.5.1 抗體輔助分析 88 4.5.2 動力學分析 90 4.5.3 小結 94 第五章 結論與未來展望 95 5.1 結論 95 5.2 未來展望 96 參考文獻 97 第六章 附錄 102 附錄一 MUC1 SEA單元候選適體挑選 102 附錄二 K2適體修飾後之結合力差異 107 附錄三 溶液離子對K2-de5’適體的結合力影響 108 附錄四 溶液離子對K2-de5’適體的結構影響 110 附錄五 溫度對K2適體的選擇性影響 112 附錄六 酸鹼值對K2 適體的結合力影響 114 附錄七 抗體與MUC1 SEA單元的結合力驗證 115 附錄八 K2適體與抗體以三明治方式於微盤分析儀測定 117 | |
| dc.language.iso | zh-TW | |
| dc.subject | 適體 | zh_TW |
| dc.subject | 表面電漿共振分析 | zh_TW |
| dc.subject | 癌症 | zh_TW |
| dc.subject | 電化學感測器 | zh_TW |
| dc.subject | 黏蛋白-1 SEA單元 | zh_TW |
| dc.subject | aptamer | en |
| dc.subject | electrochemical sensor | en |
| dc.subject | SPR analysis | en |
| dc.subject | cancers | en |
| dc.subject | Mucin-1 SEA module | en |
| dc.title | 應用適體感測黏蛋白-1 SEA單元之最佳化研究 | zh_TW |
| dc.title | Optimization of Aptamer-based Detection of Mucin-1 SEA Module | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭宗記(Hsin-Tsai Liu),周家復(Chih-Yang Tseng),魏培坤,莊旻傑 | |
| dc.subject.keyword | 癌症,黏蛋白-1 SEA單元,適體,表面電漿共振分析,電化學感測器, | zh_TW |
| dc.subject.keyword | cancers,Mucin-1 SEA module,aptamer,SPR analysis,electrochemical sensor, | en |
| dc.relation.page | 118 | |
| dc.identifier.doi | 10.6342/NTU202101697 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-08-25 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物機電工程學系 | zh_TW |
| 顯示於系所單位： | 生物機電工程學系 | |
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