PolysacDB Tetrasaccharide 3-deoxy-andalpha;-d-manno-2-octulosonic acid 1367
From DrugPedia: A Wikipedia for Drug discovery
[edit] Carbohydrate Name
Tetrasaccharide 3-deoxy-α-d-manno-2-octulosonic acid
[edit] Carbohydrate Class
Lipopolysaccharide
[edit] Source Microbe
Chlamydia spp.
[edit] Basic Structure
Consists of repeating units of tetrasaccharide of 3-deoxy--D-manno-oct-2 ulopyranosonic acid (Kdo) residues
[edit] Proposed functions
LPS represents one of the major surface antigens of this microorganism
[edit] Antigenic Nature used to produce antibodies
Glycoconjugates
[edit] Carrier Name
BSA
[edit] 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)
[edit] Antibodies
Mab S25-7
[edit] Antibody type and class
IgM
[edit] Assay System
Hemagglutination, Enzyme immunoassay, immunofluorescence, immunoblotting
[edit] Cross-reactivity
This Mab cross-reacted with elementary bodies of Chlamydia psittaci and Chlamydia trachomatis and salmonella minnesota R595 but did not cross-react with mycobacteria
[edit] Proposed epitopes
This Mab recognizes a trisaccharide epitope αKdo(2-->8) αKdo(2-->4)αKdo
[edit] Proposed Utility
This Mab can be used in the epitope characterization of Chlamydial LPS
