Rapid N-glycan release from glycoproteins using immobilized PNGase F microcolumns
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- 作者：Marton Szigetia ; Judit Bondarb ; Douglas Gjerdec ; Zsolt Keresztessyd ; Akos Szekrenyesa ; Andras Guttmana ; b ; guttman.andras@hlbs.org" class="auth_mail" title="E-mail the corresponding author
- 关键词：PNGase F ; Peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase ; GST ; glutathione-S-transferase ; RNase B ; ribonuclease B ; IgG ; immunoglobulin G ; APTS ; 8-aminopyrene-1 ; 3 ; 6-trisulfonic acid ; CGE-LIF ; capillary gel electrophoresis with laser induced fluorescence detection ; SPRI ; solid phase reversible immobilization technology ; BFS ; bare fused silica
- 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2016
- 出版时间：1 October 2016
- 年：2016
- 卷：1032
- 期：Complete
- 页码：139-143
- 全文大小：688 K
文摘
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Immobilized recombinant glutathione-S-transferase (GST) tagged PNGase F enzyme microcolumns have been developed.
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Rapid and efficient removal of N-linked carbohydrates from glycoproteins has been proved.
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The method was tested with all major N-linked glycoprotein types of: (i) neutral (IgG), (ii) highly sialylated (fetuin), and (iii) high mannose (ribonuclease B) carbohydrate containing glycoprotein standards.