Analysis of high-mannose-type oligosaccharides by microliquid chromatography-mass spectrometry and capillary electrophoresis
We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectr...
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| Published in | Electrophoresis Vol. 25; no. 13; pp. 2003 - 2009 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Weinheim
WILEY-VCH Verlag
01.07.2004
WILEY‐VCH Verlag |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0173-0835 1522-2683 |
| DOI | 10.1002/elps.200305837 |
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| Abstract | We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectrometry (ESI‐MS) and μLC/ESI‐tandem MS (MS/MS) analysis that revealed high‐mannose structure size variation between Man7GlcNAc2 and Man14GlcNAc2. Then, high‐performance CE was applied to identify possible positional isomers of the high‐mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide‐N‐glycosidase F and labeled with 1‐aminopyrene‐3,6,8‐trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man9GlcNAc2, two for Man10GlcNAc2, three for Man11GlcNAc2, Man12GlcNAc2, and Man13GlcNAc2, and two for Man14GlcNAc2. The CE results provided complementary information to the μLC/ESI‐MS and MS/MS data with respect to the possible number of positional isomers. |
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| AbstractList | We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectrometry (ESI‐MS) and μLC/ESI‐tandem MS (MS/MS) analysis that revealed high‐mannose structure size variation between Man 7 GlcNAc 2 and Man 14 GlcNAc 2 . Then, high‐performance CE was applied to identify possible positional isomers of the high‐mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide‐ N ‐glycosidase F and labeled with 1‐aminopyrene‐3,6,8‐trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man 9 GlcNAc 2 , two for Man 10 GlcNAc 2 , three for Man 11 GlcNAc 2 , Man 12 GlcNAc 2 , and Man 13 GlcNAc 2 , and two for Man 14 GlcNAc 2 . The CE results provided complementary information to the μLC/ESI‐MS and MS/MS data with respect to the possible number of positional isomers. We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectrometry (ESI‐MS) and μLC/ESI‐tandem MS (MS/MS) analysis that revealed high‐mannose structure size variation between Man₇GlcNAc₂ and Man₁₄GlcNAc₂. Then, high‐performance CE was applied to identify possible positional isomers of the high‐mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide‐N‐glycosidase F and labeled with 1‐aminopyrene‐3,6,8‐trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man₉GlcNAc₂, two for Man₁₀GlcNAc₂, three for Man₁₁GlcNAc₂, Man₁₂GlcNAc₂, and Man₁₃GlcNAc₂, and two for Man₁₄GlcNAc₂. The CE results provided complementary information to the μLC/ESI‐MS and MS/MS data with respect to the possible number of positional isomers. We report on microbore liquid chromatography (microLC) and capillary electrophoresis (CE) separation of glycopeptides and high-mannose-type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to microLC/electrospray ionization/mass spectrometry (ESI-MS) and microLC/ESI-tandem MS (MS/MS) analysis that revealed high-mannose structure size variation between Man(7)GlcNAc(2) and Man(14)GlcNAc(2). Then, high-performance CE was applied to identify possible positional isomers of the high-mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide-N-glycosidase F and labeled with 1-aminopyrene-3,6,8-trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man(9)GlcNAc(2), two for Man(10)GlcNAc(2), three for Man(11)GlcNAc(2), Man(12)GlcNAc(2), and Man(13)GlcNAc(2), and two for Man(14)GlcNAc(2). The CE results provided complementary information to the microLC/ESI-MS and MS/MS data with respect to the possible number of positional isomers. We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to μLC/electrospray ionization/mass spectrometry (ESI‐MS) and μLC/ESI‐tandem MS (MS/MS) analysis that revealed high‐mannose structure size variation between Man7GlcNAc2 and Man14GlcNAc2. Then, high‐performance CE was applied to identify possible positional isomers of the high‐mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide‐N‐glycosidase F and labeled with 1‐aminopyrene‐3,6,8‐trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man9GlcNAc2, two for Man10GlcNAc2, three for Man11GlcNAc2, Man12GlcNAc2, and Man13GlcNAc2, and two for Man14GlcNAc2. The CE results provided complementary information to the μLC/ESI‐MS and MS/MS data with respect to the possible number of positional isomers. We report on microbore liquid chromatography (microLC) and capillary electrophoresis (CE) separation of glycopeptides and high-mannose-type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to microLC/electrospray ionization/mass spectrometry (ESI-MS) and microLC/ESI-tandem MS (MS/MS) analysis that revealed high-mannose structure size variation between Man(7)GlcNAc(2) and Man(14)GlcNAc(2). Then, high-performance CE was applied to identify possible positional isomers of the high-mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide-N-glycosidase F and labeled with 1-aminopyrene-3,6,8-trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man(9)GlcNAc(2), two for Man(10)GlcNAc(2), three for Man(11)GlcNAc(2), Man(12)GlcNAc(2), and Man(13)GlcNAc(2), and two for Man(14)GlcNAc(2). The CE results provided complementary information to the microLC/ESI-MS and MS/MS data with respect to the possible number of positional isomers.We report on microbore liquid chromatography (microLC) and capillary electrophoresis (CE) separation of glycopeptides and high-mannose-type oligosaccharides, digested from recombinant phospholipase C, expressed in Pichia pastoris. The glycopeptides were subject to microLC/electrospray ionization/mass spectrometry (ESI-MS) and microLC/ESI-tandem MS (MS/MS) analysis that revealed high-mannose structure size variation between Man(7)GlcNAc(2) and Man(14)GlcNAc(2). Then, high-performance CE was applied to identify possible positional isomers of the high-mannose structures. For the CE experiments, the oligosaccharides were released from the glycoproteins by peptide-N-glycosidase F and labeled with 1-aminopyrene-3,6,8-trisulfonic acid (APTS). Excellent separation of the possible positional isomers was attained, suggesting one for Man(9)GlcNAc(2), two for Man(10)GlcNAc(2), three for Man(11)GlcNAc(2), Man(12)GlcNAc(2), and Man(13)GlcNAc(2), and two for Man(14)GlcNAc(2). The CE results provided complementary information to the microLC/ESI-MS and MS/MS data with respect to the possible number of positional isomers. |
| Author | Kreps, Joel Guttman, András Schieltz, David Koller, Antonius Khandurina, Julia Li, Jincai |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/15237400$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1016_j_jchromb_2011_08_026 crossref_primary_10_1016_j_chroma_2008_07_053 crossref_primary_10_1002_elps_200410345 crossref_primary_10_1039_C8AN01159A crossref_primary_10_1021_ac071140v crossref_primary_10_1021_ac060704c crossref_primary_10_1021_acs_analchem_6b03531 crossref_primary_10_1002_bmc_831 crossref_primary_10_1016_j_chroma_2007_04_012 crossref_primary_10_1016_j_chroma_2016_08_043 crossref_primary_10_1002_rcm_5190 crossref_primary_10_2217_fmb_09_103 crossref_primary_10_1016_j_aca_2020_08_002 crossref_primary_10_1111_1541_4337_12211 crossref_primary_10_1002_elps_200700873 crossref_primary_10_1016_j_chroma_2018_06_050 crossref_primary_10_1002_jms_799 crossref_primary_10_1002_bmc_1147 crossref_primary_10_1002_elps_201200612 crossref_primary_10_1002_bmc_1201 crossref_primary_10_1016_j_chroma_2006_12_062 crossref_primary_10_1016_j_talanta_2013_09_005 crossref_primary_10_1007_s12566_010_0018_6 |
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| Snippet | We report on microbore liquid chromatography (μLC) and capillary electrophoresis (CE) separation of glycopeptides and high‐mannose‐type oligosaccharides,... We report on microbore liquid chromatography (microLC) and capillary electrophoresis (CE) separation of glycopeptides and high-mannose-type oligosaccharides,... |
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| SubjectTerms | Capillary electrophoresis Chromatography, Liquid - methods Electrophoresis, Capillary - methods Glycopeptides Glycopeptides - analysis Glycopeptides - chemistry glycoproteins ionization Isomerism Komagataella pastoris liquid chromatography Mannose - chemistry Mass Spectrometry - methods Microbore liquid chromatography-mass spectrometry Microchemistry Oligosaccharides Oligosaccharides - analysis Oligosaccharides - chemistry Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase - metabolism phospholipase C Pichia - genetics Pichia - metabolism Pyrenes - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Time Factors Type C Phospholipases - genetics Type C Phospholipases - metabolism |
| Title | Analysis of high-mannose-type oligosaccharides by microliquid chromatography-mass spectrometry and capillary electrophoresis |
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