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Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis
Profiling of glycans released from proteins is very complex and important. To enhance the detection sensitivity, chemical derivatization is required for the analysis of carbohydrates. Due to the interference of excess reagents, a simple and reliable purification method is usually necessary for the d...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3976416/ https://www.ncbi.nlm.nih.gov/pubmed/24705408 http://dx.doi.org/10.1371/journal.pone.0094232 |
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author | Zhang, Qiwei Li, Henghui Feng, Xiaojun Liu, Bi-Feng Liu, Xin |
author_facet | Zhang, Qiwei Li, Henghui Feng, Xiaojun Liu, Bi-Feng Liu, Xin |
author_sort | Zhang, Qiwei |
collection | PubMed |
description | Profiling of glycans released from proteins is very complex and important. To enhance the detection sensitivity, chemical derivatization is required for the analysis of carbohydrates. Due to the interference of excess reagents, a simple and reliable purification method is usually necessary for the derivatized oligosaccharides. Various SPE based methods have been applied for the clean-up process. To demonstrate the differences among these methods, seven types of self-packed SPE cartridges were systematically compared in this study. The optimized conditions were determined for each type of cartridge and it was found that microcrystalline cellulose was the most appropriate SPE material for the purification of derivatized oligosaccharide. Normal phase HPLC analysis of the derivatized maltoheptaose was realized with a detection limit of 0.12 pmol (S N(−1) = 3) and a recovery over 70%. With the optimized SPE method, relative quantification analysis of N-glycans from model glycoproteins were carried out accurately and over 40 N-glycans from human serum samples were determined regardless of the isomers. Due to the high stability and sensitivity, microcrystalline cellulose cartridge showed potential applications in glycomics analysis. |
format | Online Article Text |
id | pubmed-3976416 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39764162014-04-08 Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis Zhang, Qiwei Li, Henghui Feng, Xiaojun Liu, Bi-Feng Liu, Xin PLoS One Research Article Profiling of glycans released from proteins is very complex and important. To enhance the detection sensitivity, chemical derivatization is required for the analysis of carbohydrates. Due to the interference of excess reagents, a simple and reliable purification method is usually necessary for the derivatized oligosaccharides. Various SPE based methods have been applied for the clean-up process. To demonstrate the differences among these methods, seven types of self-packed SPE cartridges were systematically compared in this study. The optimized conditions were determined for each type of cartridge and it was found that microcrystalline cellulose was the most appropriate SPE material for the purification of derivatized oligosaccharide. Normal phase HPLC analysis of the derivatized maltoheptaose was realized with a detection limit of 0.12 pmol (S N(−1) = 3) and a recovery over 70%. With the optimized SPE method, relative quantification analysis of N-glycans from model glycoproteins were carried out accurately and over 40 N-glycans from human serum samples were determined regardless of the isomers. Due to the high stability and sensitivity, microcrystalline cellulose cartridge showed potential applications in glycomics analysis. Public Library of Science 2014-04-04 /pmc/articles/PMC3976416/ /pubmed/24705408 http://dx.doi.org/10.1371/journal.pone.0094232 Text en © 2014 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhang, Qiwei Li, Henghui Feng, Xiaojun Liu, Bi-Feng Liu, Xin Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title | Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title_full | Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title_fullStr | Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title_full_unstemmed | Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title_short | Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis |
title_sort | purification of derivatized oligosaccharides by solid phase extraction for glycomic analysis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3976416/ https://www.ncbi.nlm.nih.gov/pubmed/24705408 http://dx.doi.org/10.1371/journal.pone.0094232 |
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