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| Content Provider | Springer Nature Link |
|---|---|
| Author | Blanchard, Véronique Gadkari, Rupali A. Gerwig, Gerrit J. Leeflang, Bas R. Dighe, Rajan R. Kamerling, Johannis P. |
| Copyright Year | 2006 |
| Abstract | Human chorionic gonadotropin (hCG) is a heterodimeric, placental glycoprotein hormone involved in the maintenance of the corpus luteum during the first trimester of pregnancy. Biologically active hCG has been successfully expressed in the yeast Pichia pastoris (phCG). In the context of structural studies and therapeutic applications of phCG, detailed information about its glycosylation pattern is a prerequisite. To this end N-glycans were released with peptide-N 4-(N-acetyl-β-glucosaminyl)asparagine amidase F and fractionated via anion-exchange chromatography (Resource Q) yielding both neutral (80%) and charged, phosphate-containing (20%) high-mannose-type structures. Subfractionations were carried out via normal phase (Lichrosorb-NH2) and high-pH anion-exchange (CarboPac PA-1) chromatography. Structural analyses of the released N-glycans were carried out by using HPLC profiling of fluorescent 2-aminobenzamide derivatives, MALDI-TOF mass spectrometry, and 500-MHz 1H-NMR spectroscopy. Detailed neutral oligosaccharide structures, in the range of Man8GlcNAc2 to Man11GlcNAc2 including molecular isomers, could be established, and structures up to Man15GlcNAc2 were indicated. Phosphate-containing oligosaccharides ranged from Man9 PGlcNAc2 to Man13 PGlcNAc2. Mannosyl O-glycans were not detected. Profiling studies carried out on different production batches showed that the oligosaccharide structures are similar, but their relative amounts varied with the culturing media. |
| Starting Page | 33 |
| Ending Page | 47 |
| Page Count | 15 |
| File Format | |
| ISSN | 02820080 |
| Journal | Glycoconjugate Journal |
| Volume Number | 24 |
| Issue Number | 1 |
| e-ISSN | 15734986 |
| Language | English |
| Publisher | Kluwer Academic Publishers-Plenum Publishers |
| Publisher Date | 2006-12-05 |
| Publisher Place | New York |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Glycosylation Pichia pastoris Human chorionic gonadotropin High-mannose-type N-glycans Phosphorylation Pathology Biochemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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