| PolysacDB ID | 2611 |
| Carbohydrate Name | Glucurunoxylomannan (Drugpedia) |
| Carbohydrate Class | Capsular polysaccharide |
| Microbe | Cryptococcus neoformans serotype A (NCBI Taxonomy) (Drugpedia) |
| Basic Structure | The capsular polysaccharide has a linear α-(1-->3)-linked mannose backbone that is substituted with nonreducing D-xylosyl and D-glucuronosyl acid groups. The mannose backbone is partially O-acetylated
|
| BCSDB Structure | 116826 |
| Proposed functions | The capsular polysaccharide is an important virulence factor |
| Antigenic Nature used to produce antibodies | Glycoconjugates |
| Carrier Name | Tetanus toxoid |
| Conjugation Method | GXM was derivatized by the following two methods. (i) In method 1, ADH [adipic acid dihydrazide] was introduced into GXM by the activation of carboxyl groups with EDAC. GXM (5 mg/ml of 0.2 M NaCl) was derivatized with 0.5 M ADH and 0.1 M EDAC at pH 4.85 for 3.5 h at room temperature, using a pH Stat. After extensive dialysis against 0.2 M NaCl, the reaction mixture was passed through a 2B-CL Sepharose column (1.5 by 30 cm) equilibrated in 0.2 M NaCl. The fractions containing GXM were pooled and concentrated to the original volume. (ii) In method 2, ADH was introduced into GXM by the activation of hydroxyl groups with CNBr. GXM (5 mg/ml of 0.2 M NaCl) was activated with an equal weight of CNBr at pH 10.5 for 6 min at 4°C, using a pH Stat. An equal volume of 0.5 M NaHCO3 (pH 8.5) containing 0.5 M ADH was added. The reaction mixture was tumbled at 3 to 8°C for 18 to 20 h, dialyzed against 0.2 M NaCl, and passed through a 2B-CL Sepharose column (1.5 by 30 cm). The fractions containing GXM were pooled and concentrated to the original volume. The reaction mixture, containing equal concentrations (3.0 to 7.5 mg/ml) of GXM-AH (derivatized by either method) and TT or rEPA in 0.2 M NaCl, was brought to pH 5.6 with 0.05 N HCl, and 0.05 to 0.1 M EDAC was added; the pH was maintained at 5.6 in a pH Stat for 1 to 3 h at 4°C. The reaction mixture was dialyzed against 0.2 M NaCl at 3 to 8°C and passed through a Sepharose 2B-CL column (1.5 by 30 cm) equilibrated in 0.2 M NaCl. The void volume fractions containing the GXM and the protein were pooled and stored in 0.01% thimerosal at 3 to 8°C |
| Antibodies | Mab 2H1 |
| Antibody type and class | IgG1 |
| Assay System | ELISA |
| Cross-reactivity | This antibody was cross-reactive to serotypes A,B,C and D |
| Proposed epitopes | This antibody did not bind O-deacetylated glucurunoxylomannan indicating that acetylated residues on glucurunoxylomannan surface are an essential part of the epitope |
| IEDB Epitope | 115576 |
| Proposed Utility | Could help in specific serotype identification |
| Curator ID | AA + AS |
| Date of Curation | 17-07-2011
|
| References | 1583327 |