跳到主要內容

臺灣博碩士論文加值系統

(3.231.230.177) 您好!臺灣時間:2021/07/28 23:13
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:范文欽
研究生(外文):Wen-Chin Fan
論文名稱:探討蛋白質PEGylation的反應特性及其應用在重組腺病毒的修飾
論文名稱(外文):Characterization of Protein PEGylation and its Application on Modification of Recombinant Adenovirus
指導教授:徐祖安
指導教授(外文):Tsu-An Hsu
學位類別:碩士
校院名稱:國立清華大學
系所名稱:化學工程學系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:57
中文關鍵詞:腺病毒
外文關鍵詞:adenovirusPEGylationstability
相關次數:
  • 被引用被引用:0
  • 點閱點閱:215
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
我們利用Tresyl monomethoxy polyethylene glycol (TM-PEG) 與free lysine的PEGylation反應,探討其動力學模式,利用螢光分析及作圖法,估計出TM-PEG及Lysine於PEGylation反應中的反應級數分別為1.69及1.09,同時並以Arrhenius學說為基礎,推論PEGylation反應的活化能,進而探討反應對溫度的敏感性。

在了解反應物濃度、溫度及時間對PEGylation反應的影響後,將此結果應用於重組腺病毒的PEGylation反應,藉由提昇反應物濃度,以降低反應的時間,同時配合添加少許的蔗糖 (5 %),可以改善因PEGylation反應造成病毒活性大幅流失的現象。雖然,經過PEGylation反應後的腺病毒依然有活性的流失,但PEGylation卻提昇了病毒的穩定性,根據結果顯示,在37 ℃環境下平均約每小時流失1個 log 值的活性,降低至約每小時 0.5-1個log 值的活性流失。而在4 ℃的溫度下,由一天1.5 log 值左右的喪失活性,減少至一天 1 個 log值以內的活性喪失。
We studied the kinetics of PEGylation by observing the conjugation reaction between tresyl monomethoxy polyethylene glycol (TM-PEG) and free lysine. We employed a fluorescence assay and graphic methods to determine the reaction orders with respective to TM-PEG and lysine. According to the experimental results, the reaction orders of PEG and lysine are 1.69 and 1.09, respectively. We also determined the activation energy of PEGylation using Arrhenius’ theory and discussed the temperature sensitivity of PEGylation.

The reaction kinetics was applied to optimize that PEGylation reaction to modify adenovirus. By raising reactant concentrations and with the addition of 5 ﹪sucrose in the reaction mixture, the loss of adenoviruses titer during PEGylation process could be reduced. Although adenovirus titer was reduced during PEGylation, PEGylated adenovirus exhibits better stability under harsh storage conditions. Our results showed that there was approximately 0.5 to 1 log loss of the original titer in PEGylated adenovirus per hour at 37℃ whereas there was at least 1 log per hour reduction in viral titer when non-PEGylated adenovius was incubated at 37℃. Similarly, PEGylated adenovirus is more stable than non-PEGylated adenovirus at 4 ℃; the rate of titer loss is 1.5 log/day and 1.0 log/day for non-PEGylated and PEGylated adenoviruses, respectively.
1. Abuchowski, A., J. R. McCoy, N. C. Palczuk, v. E. T, and F. F. Davis. 1977. Effect of covalent attachment of polyethylene glycol on immunogenicity and circulating life of bovine liver catalase. J Biol Chem 252:3582-3586.
2. Abuchowski, A., v. E. T., P. N. C., and D. F.F. 1977. Alteration of immunogical properties of polyethylene glycol. J. Biol. Chem 252:3578-3581.
3. Andrew, P., C. 2002. PEGylated antibodies and antibody fragments for improved therapy: a review. Advanced Drug Delivery Reviews 54:531-545.
4. Armentan, D., J. Zabner, C. Sacks, C. C. Sookdeo, P. Smith M, J. A. George, S. C. Wadsworth, A. E. Smith, and R. J. Gregory. 1997. Effect of the E4 region on the persistence of transgene expression from adenovirus vectors. J. Virol. 71:2408-2416.
5. Bai, M., B. Harfe, and P. Freimuth. 1993. Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells. Journal of Virology 67:5198-5205.
6. Benihoud, K., P. Yeh, and M. Perricaudet. 1999. Adenovirus vectors for gene therapy. Current Opinion in Biotechnology 10:440-447.
7. Bennett, E., J. R. Bennink, J. W. Yewdell, and F. M. Brodsky. 1999. Cutting edge : adenovirus E19 has two mechanisms for class 1 MHC expression. The Journal of Immunology 162:5049-5052.
8. Bergelson, J. M., J. A. Cunningham, G. Droguett, E. A. Kurt-Jones, A. Krithivas, J. S. Hong, M. S. Horwitz, R. L. Crowell, and R. W. Finberg. 1997. Isolation of common receptor for coxsackie B viruses and adenoviruses 2 and 5. Science 275:1320-1323.
9. Cheng, T. L., P. Y. Wu, M. F. Wu, J. W. Chern, and S. R. Roffler. 1999. Accelerated clearance of polyethylene glycol-modified proteins by anti-polyethylene glycol IgM. Bioconjugate Chemistry 10:520-528.
10. Croyle, M. A., X. Cheng, and J. M. Wilson. 2001. Development of formulations that enhance physical stability of viral vectors for gene therapy. Gene Therapy 8:1281-1290.
11. Croyle, M. A., N. Chirmule, Y. Zhang, and J. M. Wilson. 2002. PEGylation of E1-deleted adenovirus vectors allows significant gene expression on readministration to liver. Human Gene Therapy 13:1887-1900.
12. Croyle, M. A., B. J. Roessler, B. L. Davidson, J. M. Hilfinger, and G. L. Amidon. 1998. Factors that influence stability of recombinant adenoviral preparations for human gene therapy. Pharmaceutical Development and Technology 3:373-383.
13. Croyle, M. A., Q. C. Yu, and J. M. Wilson. 2000. Development of a rapid method for the PEGylation of adenovirus with enhanced transduction and improved stability under harsh storage conditions. Human Gene Therapy 11:1713-1722.
14. Delgado, C., G. Francis, and D. Fisher. 1992. The usesand properties of PEG-linked proteins. Crit. Rev. Ther. Drug Carrier Syst 9:249-304.
15. Delgado, C., J. N. Patel, G. E. Francis, and D. Fisher. 1990. Coupling of poly(ethylene glycol) to albumin under very mild conditions by activation with trsyl chloride: characterization of the conjugate by partitioning in aqueous two-phase systems. Biotechnology and Applied Biochemistry 12:119-128.
16. Delgado, M., F. Malik, B. FSelisko, D. Fisher, and G. E. Francis. 1994. Quantitative analysis of polyethylene glycol(PEG) in PEG-modified proteins/cytokines by aqueous two-phase system. Journal of Biochemical and Biophysical Methods 29:237-250.
17. Enders, J., J. Bell, and J. Dingle. 1956. ''Adenoviruses'':group name proposed for new respiratory -tract viruses. science 124:119-120.
18. Fields, B. N., D. M. knipe, and P. M. Howley. 1995. Fields Virology, vol. 2.
19. Gao, G. P., Y. Yang, and J. M. Wilson. 1996. Biology of adenovirus vectors with E1 and E4 deletions for liver-directed gene therapy. Journal of Virology 70:8934-8943.
20. Graham, F. L., and L. Prevec. 1992. Adenovirus-based expression vectors and recombinant vaccines. biotechnology 20:363-390.
21. Graham, F. L., J. Smiley, W. C. Russell, and R. Nairn. 1977. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. Journal General Virology 36:59-72.
22. Greber, U. F., M. Willetts, P. Webster, and A. Helenius. 1993. Stepwise dismantling of adenovirus 2 during entry into cells. Cell 75:477-486.
23. Han, J., P. Sabbatini, L. Rao, D. Modha, and E. White. 1996. The E1B 19K protein blocks apoptosis by interacting with and inhibiting the p53-inducible and deth-promoting Bax protein. Gene and Develop 10:461-477.
24. Hosakawa, K., and M. T. Sung. 1976. Isolation and characterization of an extremely basic protein from adenovirus type 5. Journal of Virology 17:924-934.
25. Kannabiran, C., G. F. Morris, and M. B. Mathews. 1999. Dual action of the adenovirus E1A 234R oncoprotein on the human proliferating cell nuclear antigen promoter : repression of transcriptional activation by p53. Oncogene 18:7825-7833.
26. Kenneth, D. H., and S. W. Kim. 2002. Effects of PEG conjugation on insulin properties. Advanced Drug Delivery Reviews 54:505-530.
27. Lai, C. M., K. Y. Lai, and P. E. Rakoczy. 2002. Adenovirus and adeno-associated virus vectors. DNA and Cell Biology 21:895-913.
28. Lusky, M. 1998. In vitro and in vivo biology of recombinant adenovirus vectors with E1, E1/E2A, or E1/E4 deleted. Journal of Virology 72:2022-2032.
29. Milton Harris, J., N. E. Nancy, and M. Marlene. 2001. Pegylation: A Novel Process for modifying Pharmacokinrtics. Clinical Pharmacokinetics 40:539-551.
30. Nunes, F. A., E. E. Furth, J. M. Wilson, and S. E. Raper. 1999. Gene transfer into the liver of non-human primates with E1-deleted recombinant adenoviral vectors: Safety of re-administration. 10. 2515-2526. Human Gene Therapy 10:2515-2526.
31. O'Riordan, C. R., A. Lachapelle, C. Delgado, V. Parkes, S. C. Wadsworth, A. E. Smith, and G. E. Francis. 1999. PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo. Human Gene Therapy 10:1349-1358.
32. Parks, R. J., C. M. Evelegh, and F. L. Graham. 1996. Use of helper-dependent adenoviral vectors of alternative serotypes permits repeat vector administration. Gene Therapy 6:1565-1573.
33. Pettersson, U., and R. J. Roberts. 1986. Adenovirus gene expression and replication : a historical review. Cancer cells 4:37-57.
34. Romanczuk, H., C. E. Galer, J. Zabner, G. Barsomian, S. C. Wadsworth, and C. R. O'Riordan. 1999. Modification of an adenoviral vector with biologically selected peptides: a novel strategy for gene delivery to cells of choice. Human Gene Therapy 10:2615-2626.
35. Rowe, W. P., R. J. Huebner, and L. K. Gilmore. 1953. Isolation of a cytopathogenic agent from human adenoids undergoing spontaneous degeneration in tissue culture. Proceedings of the Society for Experimental Biology and Medicine 65:570-573.
36. Russell, W. C. 2000. Update on adenovirus and its vectors. Journal of General. Virology 81:2573-2604.
37. Russell, W. C., W. G. Laver, and P. J. Sanderson. 1968. Internal components of adenovirus. nature 219:1127-1130.
38. Shabram, P. W., D. D. Giroux, A. M. Goudreau, R. J. Gregory, M. T. Horn, B. G. Huyghe, X. Liu, M. H. Nunnally, B. J. Sugarman, and S. Sutjipto. 1997. Analytical anion-exchange HPLC of recombinant type-5 andenovirus particles. Human Gene Therapy 8:453-456.
39. Stephan, A., Vorburger, K. Kelly, and Hunt. 2002. Adenoviral Gene Therapy. The Oncologist 7:46-59.
40. Stewart, P. L., R. M. Burnett, M. Cyrklaff, and S. D. Fuller. 1991. Image reconstruction reveals the complex molecular organization of adenovirus. cell 67:145-154.
41. Stewart, P. L., S. D. Fuller, and R. M. Burnett. 1993. Difference imaging of adenovirus: bridging the resolution gap between X-ray crystallography and electron microscopy. The EMBO Journal 7:2589-2599.
42. Tomko, R. P. X. R., and L. Philipson. 1997. HCAR and MAR: The human and mouse cellular receptors for subgroup C adenoviruses and group B coxsackieviruses. Proceedings of the National Academy of Sciences of the United States of America 94:3352-3356.
43. Tsujinoue, H., S. Kuriyama, K. Tominaga, H. Okuda, T. Nakatani, H. Yoshiji, T. Tsujimoto, T. Akahane, K. Asada, and H. and Fukui. 2001. Intravenous re-administration of an adenoviral vector performed long after the initial administration failed to induce re-expression of the original transgene in rats. International Journal of Radiation Oncology, Biology, Physics 18:575-580.
44. Valentine, R. G., and H. G. Pereira. 1965. Antigens and structure of the adenovirus. Journal of Molecular Biology 13:13-20.
45. Vorburger, S. A., and K. K. Hunt. 2002. Adenoviral gene therapy. The Oncologist 7:46-59.
46. Wang, K., T. Guan, D. A. Cheresh, and G. R. Nemerow. 2000. Regulation of adenovirus membrane penetration by the cytoplasmic tail of integrin ?. Journal of Virology 74:2731-2739.
47. Wickham, T. J., P. Mathias, D. A. Cheresh, and G. R. Nemerow. 1993. Intergrins alpha3 and alpha5 promote adenovirus internalization but not virus attachment. Cell 73:309-331.
48. Worgall, S., G. Wolfe, E. Falck-Pedersen, and R. G. Crystal. 1997. Innate immune mechanisms dominate elimination of adenoviral vectors following in vivo administration. Human Gene Therapy 8:37-44.
49. Yang, Y., H. C. J. Ertl, and J. M. Wilson. 1994. MHC class Irestricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses. Immunity 1:433-442.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top