Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation
碩士 === 國立嘉義大學 === 應用化學系研究所 === 101 === Peptides of digested protein are not fully observed by MS-based proteomic methods, such as LC-ESI-MS. A peptide was classified as proteotypic when the number of observation is 100% of all injection numbers. Increasing the number of peoteotypic peptides will the...
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ndltd-TW-101NCYU55000242016-03-18T04:41:38Z http://ndltd.ncl.edu.tw/handle/96077867680488158156 Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation 胜肽性質及其在 LC-ESI-MS 偵測之研究 Shih-Ying Tai 戴詩穎 碩士 國立嘉義大學 應用化學系研究所 101 Peptides of digested protein are not fully observed by MS-based proteomic methods, such as LC-ESI-MS. A peptide was classified as proteotypic when the number of observation is 100% of all injection numbers. Increasing the number of peoteotypic peptides will therefore give more accurate information for protein identification. For the detected number of peptides, electrospray ionization efficiency has a great influence. And the factor divide into the analyte nature and instrument parameters. In the present study, we aim at discuss analyte property in fixed LC-ESI-MS parameters conditions. However, for the separation of peptides of different polarity, different analytic solvent environment could cause different ionization efficiency. Therefore, the amount of the proteotypic peptides detected in ESI-MS is usually restricted by the low ionization efficiency of those highly water soluble peptides presented in the digested mixture. Here, we provided a method, post-column solvent compensated device in LC-ESI-MS for increasing the detection number of proteotypic peptides via adjust the gradient LC eluents to a specific relative nonpolar solvent composition. In order to maximize the ionization efficiency for peptides with various polarities, the ratio of make-up solvent based on authentic peptides is 50% MeOH. The method is demonstrated by uthentic peptides (CALNNPKEK, LSA and GLGLG) and enzymatic digested model proteins (BSA, ovalbumin, and β-lactoglobulin). The MS peak area of CALNNPKEK detected under make-up condition (make-up to 50% MeOH) was 2.04 times to that of the non-make-up condition. And the followed species LSA and GLGLG were 1.65 and 1.26 times to the non-make-up ones, respectively. Those data revealed that the ionization efficiencies of authentic peptides in LC-ESI-MS were all improved. The detected number of proteotypic peptides of the enzymatic digested model proteins increased by the polarity of the original proteins. Kuo-Lung Ku 古國隆 學位論文 ; thesis 93 zh-TW |
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碩士 === 國立嘉義大學 === 應用化學系研究所 === 101 === Peptides of digested protein are not fully observed by MS-based proteomic methods, such as LC-ESI-MS. A peptide was classified as proteotypic when the number of observation is 100% of all injection numbers. Increasing the number of peoteotypic peptides will therefore give more accurate information for protein identification. For the detected number of peptides, electrospray ionization efficiency has a great influence. And the factor divide into the analyte nature and instrument parameters. In the present study, we aim at discuss analyte property in fixed LC-ESI-MS parameters conditions. However, for the separation of peptides of different polarity, different analytic solvent environment could cause different ionization efficiency. Therefore, the amount of the proteotypic peptides detected in ESI-MS is usually restricted by the low ionization efficiency of those highly water soluble peptides presented in the digested mixture. Here, we provided a method, post-column solvent compensated device in LC-ESI-MS for increasing the detection number of proteotypic peptides via adjust the gradient LC eluents to a specific relative nonpolar solvent composition. In order to maximize the ionization efficiency for peptides with various polarities, the ratio of make-up solvent based on authentic peptides is 50% MeOH. The method is demonstrated by uthentic peptides (CALNNPKEK, LSA and GLGLG) and enzymatic digested model proteins (BSA, ovalbumin, and β-lactoglobulin). The MS peak area of CALNNPKEK detected under make-up condition (make-up to 50% MeOH) was 2.04 times to that of the non-make-up condition. And the followed species LSA and GLGLG were 1.65 and 1.26 times to the non-make-up ones, respectively. Those data revealed that the ionization efficiencies of authentic peptides in LC-ESI-MS were all improved. The detected number of proteotypic peptides of the enzymatic digested model proteins increased by the polarity of the original proteins.
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author2 |
Kuo-Lung Ku |
author_facet |
Kuo-Lung Ku Shih-Ying Tai 戴詩穎 |
author |
Shih-Ying Tai 戴詩穎 |
spellingShingle |
Shih-Ying Tai 戴詩穎 Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
author_sort |
Shih-Ying Tai |
title |
Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
title_short |
Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
title_full |
Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
title_fullStr |
Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
title_full_unstemmed |
Improvement of peptide detection by LC-ESI-MS;Focus on solvent compensation |
title_sort |
improvement of peptide detection by lc-esi-ms;focus on solvent compensation |
url |
http://ndltd.ncl.edu.tw/handle/96077867680488158156 |
work_keys_str_mv |
AT shihyingtai improvementofpeptidedetectionbylcesimsfocusonsolventcompensation AT dàishīyǐng improvementofpeptidedetectionbylcesimsfocusonsolventcompensation AT shihyingtai shèngtàixìngzhìjíqízàilcesimszhēncèzhīyánjiū AT dàishīyǐng shèngtàixìngzhìjíqízàilcesimszhēncèzhīyánjiū |
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