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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 翁啟惠 | zh_TW |
| dc.contributor.advisor | Chi-Huey Wong | en |
| dc.contributor.author | 王寶源 | zh_TW |
| dc.contributor.author | Pao-Yuan Wang | en |
| dc.date.accessioned | 2024-03-07T16:27:28Z | - |
| dc.date.available | 2024-03-08 | - |
| dc.date.copyright | 2024-03-07 | - |
| dc.date.issued | 2016 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | 1. Fuster MM & Esko JD (2005) The sweet and sour of cancer: glycans as novel therapeutic targets. Nature reviews. Cancer 5(7):526-542.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/92180 | - |
| dc.description.abstract | zh_TW | |
| dc.description.abstract | Mass-spectrometry (MS)-based glycomics aims to map all the molecular glycan species under specific physiological or pathological conditions. Glycosphingolipids (GSLs) are found to be involved in many biological processes. Overexpression of GSLs in different cancers has been observed widely, including in glioblastoma multiforme (GBM). The conventional MS method used for GSLs detection is mostly MALDI. However, the poor capacity for separation of isomeric molecules of MALDI is not sufficient for desired complete glycomic mapping. In this thesis, we developed a porous graphitized carbon (PGC)-LC-MS based workflow to systematically analyze all glycans released from GSLs in cells. By using acidic solvent system in positive mode, six GM1 (Hex3HexNAc1NeuAc1) standards were mixed together and then loaded onto PGC-LC-MS for separation. Under optimized gradient condition, the six standards were completely separated and their structures were also confirmed by CID-MS/MS. After platform being established, GSLs glycans of DBTRG GBM cells were applied to this platform for the isomer separation. The GM1 isomers (Hex3HexNAc1NeuAc) of DBTRG cells were separated, identified and quantified for the first time. Moreover, the whole GSLs of DBTRG cells could be separated and analyzed on the PGC-LC-MS system. Minor species, such as globo-series molecules, were also detected and several other isomeric glycans were identified. Next, this method was used to compare the changed expression of GSLs between DBTRG parental cells and stem-like neurospheres. The increased level of sialylated and decreased level of neutral GSLs were detected in neurospheres. Finally, the platform was further applied to profile and compare the intact GSLs of other GBM cells. The short GSLs usually have fewer isomers because of the simple molecular structures. On the other side, long GSLs usually have more isomeric ions because of their complicated structures. There are some unusual isomeric glycans being detected, such as Gb5/iGb5 distribution in each kind of cells examined. In short, we here developed a useful PGC-LC-MS platform for isomeric GSLs separation by LC and identification by MS/MS and it can be applied to more biological samples in the future. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-03-07T16:27:28Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2024-03-07T16:27:28Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | Chapter 1: Introduction 1
1.1 Glycosylation 1 1.2 Glycosphingolipids (GSLs) and its Biological Importance 4 1.3 Glycosphingolipids (GSLs) and Glioblastoma Multiforme (GBM) 9 1.4 Mass spectrometry (MS)-Based Separation of Isomeric Glycosphingolipids 13 1.5 Specific Aims 17 Chapter 2: Materials and Methods 19 2.1 Chemicals and Reagents 19 2.2 Cell Culture 19 2.3 Extraction of Glycosphingolipids 20 2.4 Permethylation 20 2.5 MALDI-MS and MS/MS 21 2.6 nanoLC-ESI-PGC-MS and MS/MS 21 Chapter 3: Results 23 3.1 Establishment of PGC-LC-MS/MS-based GSLs profiling and isomeric analysis 23 3.1.1 MALDI-MS and MS/MS analysis of GSLs of DBTRG cells 23 3.1.2 Nano-Porous Graphitized Carbon (PGC)-LC-MS and MS/MS analysis of GSLs glycans 28 3.1.3 Discussion 35 3.2 Comparison of GSLs profiles between DBTRG and DBTRG neurospheres 38 3.2.1 MALDI-MS profiles of DBTRG and DBTRG neurospheres 38 3.2.2 PGC-LC-MS and MS/MS of DBTRG and DBTRG neurospheres 39 3.2.3 Discussion 41 3.3 Isomeric analysis of GSLs of other GBM cells 42 3.3.1 PGC-LC-MS and MS/MS of LN229 cells 42 3.3.2 PGC-LC-MS and MS/MS of Hs683 cells 44 3.3.3 PGC-LC-MS and MS/MS of U87 cells 45 3.3.4 Discussion 46 Chapter 4: Discussion and Conclusion 48 4.1 Current status of MS-based glycan isomer separation 48 4.2 Future perspectives for MS-based glycan isomer separation 51 4.3 Biological implications from MS-based glycan isomer separation 53 References 54 | - |
| dc.language.iso | en | - |
| dc.subject | 孔洞石墨碳管柱 | zh_TW |
| dc.subject | 神經膠質母細胞瘤 | zh_TW |
| dc.subject | 醣脂體 | zh_TW |
| dc.subject | Porous Graphitized Carbon (PGC) | en |
| dc.subject | Glycosphingolipids | en |
| dc.subject | Glioblastoma | en |
| dc.title | 孔洞石墨碳管柱液相層析串連式質譜分析技術之研發與其在分離與結構鑑定神經膠質母細胞瘤之醣脂體醣鏈之應用 | zh_TW |
| dc.title | Development of Porous Graphitized Carbon (PGC)-LC-MS/MS for the Separation and Structural Determination of the Glycans from Glycosphingolipids in Glioblastoma cells | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 104-2 | - |
| dc.description.degree | 博士 | - |
| dc.contributor.oralexamcommittee | 邱繼輝;林俊宏;林國儀;沈家寧 | zh_TW |
| dc.contributor.oralexamcommittee | ;;; | en |
| dc.subject.keyword | 孔洞石墨碳管柱,神經膠質母細胞瘤,醣脂體, | zh_TW |
| dc.subject.keyword | Porous Graphitized Carbon (PGC),Glycosphingolipids,Glioblastoma, | en |
| dc.relation.page | 102 | - |
| dc.identifier.doi | 10.6342/NTU201602023 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2016-08-09 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生化科學研究所 | - |
| 顯示於系所單位： | 生化科學研究所 | |
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