| Autor: |
Gaurang P Bhide, Natalie E Bohrer, Danying Song, Jeanne Cabel, Marc R Lake, John E Harlan, Laura J Miesbauer |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Glycobiology. |
| ISSN: |
1460-2423 |
| Popis: |
Monoclonal antibodies comprise a major class of biologic therapeutics and are also extensively studied in immunology. Given the importance of glycans on antibodies, fluorescent labeling of enzymatically released glycans and their LC/MS analysis is routinely used for in-depth characterization of antibody glycosylation. In this technical note, we propose a method for facile characterization of glycans in the variable region of antibodies using sequential enzymatic digests with Endoglycosidase-S2 and RapidTM Peptide-N-Glycosidase-F followed by labeling with a fluorescent dye carrying an NHS-carbamate moiety. The results and proposed mechanism also suggest that the choice of glycosidases along with the labeling chemistry is critical for accurate glycan analysis for a desired application. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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