| Chapter title |
Preparation of Glycan Arrays Using Pyridylaminated Glycans
|
|---|---|
| Chapter number | 16 |
| Book title |
Microarray Technology
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3136-1_16 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3135-4, 978-1-4939-3136-1
|
| Authors |
Shin-ichi Nakakita, Jun Hirabayashi |
| Abstract |
We describe the method to prepare neoglycoproteins from the conjugation of bovine serum albumin and pyridylaminated glycans. Large quantities of glycans (>1 mg) can be pyridylaminated and then converted to their 1-amino-1-deoxy derivatives by reaction with hydrogen followed by hydrazine. These pyridylaminated glycans can then be conjugated to bovine serum albumin via esterification with N-( m-maleimidobenzoyloxy)succinimide to form a neoglycoprotein, e.g., glycosylated bovine serum albumin. As a demonstration, we prepared High-mannose bovine serum albumin, which was immobilized on an activated glass slide. Then, we showed that the neoglycoprotein bind to Cy3-labeled Lens culinaris agglutinin, a mannose-specific plant lectin, as detected using an evanescent-field-activated fluorescence scanner system. |
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