Immobilisation of antibodies in gels allows the improved release and identification of glycans
| Autor: | Patrick Camilleri, J. Mark Skehel, Joanne Charlwood |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Glycan
Proteome Molecular Sequence Data Mass spectrometry Biochemistry High-performance liquid chromatography Antibodies Immunoglobulin G chemistry.chemical_compound Polysaccharides Animals Humans Sodium dodecyl sulfate Molecular Biology Polyacrylamide gel electrophoresis Chromatography High Pressure Liquid Fluorescent Dyes chemistry.chemical_classification Chromatography biology Chemistry Hydrophilic interaction chromatography Antibodies Monoclonal Sodium Dodecyl Sulfate Carbohydrate Sequence Immunoglobulin M Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Cattle Electrophoresis Polyacrylamide Gel Glycoprotein Gels |
| Zdroj: | PROTEOMICS. 1:275-284 |
| ISSN: | 1615-9861 1615-9853 |
| DOI: | 10.1002/1615-9861(200102)1:2<275::aid-prot275>3.0.co;2-k |
| Popis: | Human IgG and IgM, bovine IgM and three therapeutic IgG monoclonal antibodies have been separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Carbohydrates were then released from these immobilised proteins by direct enzymatic digestion, derivatised with a highly fluorescent probe and analysed by high performance liquid chromatography and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. This procedure not only allowed measurement of the purity of the intact antibodies but also provided detailed analysis of the complex mixtures of oligosaccharides covalently attached to these glycoproteins. The methodology out-lined allows the simultaneous processing of a number of glycoproteins separated on one single gel. In contrast to the release of carbohydrate from glycoproteins in solution, this procedure can also be conveniently applied when only impure glycoprotein is available. |
| Databáze: | OpenAIRE |
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