| PolysacDB ID | 1369 |
| Carbohydrate Name | Tetrasaccharide 3-deoxy-α-d-manno-2-octulosonic acid (Drugpedia) |
| Carbohydrate Class | Lipopolysaccharide |
| Microbe | Chlamydia spp. (NCBI Taxonomy) (Drugpedia) |
| Basic Structure | Consists of repeating units of tetrasaccharide of 3-deoxy--D-manno-oct-2 ulopyranosonic acid (Kdo) residues
|
| BCSDB Structure | N/A |
| Proposed functions | LPS represents one of the major surface antigens of this microorganism |
| Antigenic Nature used to produce antibodies | Glycoconjugates |
| Carrier Name | Bovine serum albumin |
| Conjugation Method | Allylglycosides were reacted with cysteamine leading to ligands with spacers of the general formula R-(CH2)3-S-(CH2)2-NH3+, where R represents the respective carbohydrate ligand. These compounds were reacted with acryloyl chloride, resulting in the formation of derivatives of the general formula R-(CH2)3-S-(CH2)2- NH-CO-CH=CH2, which, after copolymerization with acrylamide, yielded polyacrylamide derivatives. Ligands with spacers of the general formula R-(CH2)3-S-(CH2)2-NH3' were activated into isothiocyanate derivatives as follows. Thiophosgene (6 ul) was dissolved in chloroform (3 ml), and a solution of the ligand with spacers (15 mg) in water (3 ml) was added. The reaction was monitored by using thin-layer chromatography on plates of silica gel 60 with chloroformmethanol-water (10/10/3 by volume) as the irrigant and detection with UV light (254-nm wavelength). When the reaction was completed, usually within 3 h of stirring at room temperature, the organic phase was removed and the aqueous layer was extracted three times with 3 ml of chloroform. The aqueous phase was freed from residual chloroform by evaporation under a stream of nitrogen and added to a solution of BSA in 0.3 M sodium chloride containing 0.1 M sodium bicarbonate (1 ml). The reaction mixture was stirred for 48 h at room temperature and then separated by gel permeation chromatography with a 35- by 1.6-cm column of Sephadex G-50 and 10 mM sodium bicarbonate as the eluant. Collected ninhydrin-positive fractions were combined and dialyzed against phosphate-buffered saline (0.15 M, pH 7.2; PBS) |
| Antibodies | Mab S25-2 |
| Antibody type and class | IgG1 |
| Assay System | Hemagglutination, Enzyme immunoassay, immunofluorescence, immunoblotting |
| Cross-reactivity | This Mab cross-reacted with elementary bodies of Chlamydia psittaci and Chlamydia trachomatis but did not cross-react with mycobacteria and and salmonella minnesota R595 |
| Proposed epitopes | This Mab recognizes a trisaccharide epitope αKdo(2-->8) αKdo(2-->4)αKdo |
| IEDB Epitope | 76828 |
| Proposed Utility | This Mab can be used in the epitope characterization of Chlamydial LPS |
| Curator ID | AA + AS |
| Date of Curation | 30-06-2010
|
| References | PMC256998 |