CarboDB Capsular polysaccharide 1005
From DrugPedia: A Wikipedia for Drug discovery
Revision as of 00:00, 15 January 2001
Carbohydrate Name
Capsular polysaccharide
Carbohydrate Class
Capsular polysaccharide
Source Microbe
Haemophilus influenzae
Basic Structure
A relatively simple antigen consisting of repeating units of 3-β-D ribose-(1-->1)-D-ribitol-5-phosphate
Proposed functions
An important virulence determinant
Antigenic Nature used to produce antibodies
Glycoconjugates
Carrier Name
Tetanus toxoid
Conjugation Method
Protein solutions (25 mg/ml) and Adipic acid dihydrazide [ADH] (3.45 rag/rag protein) were reacted with three different concentrations of EDAC (0 1, 0.3, and 0.6 rag/rag protein) The pH of the reaction mixture was maintained at 4.7 ± 0.2 with 0.1 N HCL. The reaction proceeded at room temperature for 3 h and the reaction mixtures were dialyzed at 3-8C with two changes/d against 6 liter of 0 2 M NaCI. The albumin and polysaccharide derivatives were then dialyzed against two 6-liter changes of deionized water and freeze-dried. The polysaccharide was activated with CNBr. Briefly, a solution of polysaccharide (5.0 mg/ml), equilibrated at 4C, was rapidly brought to pH 10.5 with 0.1 N NaOH. 100 mg/ml CNBr was added to a final concentration of 0.4 mg/mg polysaccharide, and the pH maintained at 10.5 for 6 rain. Then the reaction mixture was brought to pH 8 5 with 0.5 M NaHCO3, and the CNBr-actlvated polysaccharide added to an equal weight of ADH-protein. The reaction mixture was tumbled gently overnight at 3-8C and then centrifuged at 16,000 g, 4C for 20 ram. The supernatant was passed through a CL-4B Sepharose column, 1 5 × 90 cm, that was equihbrated with 0 2 M ammonium acetate. The void-volume fractions were pooled, dialyzed against 0.01 M phosphate-buffered 0.145 M NaCI, pH 7.0, at 3-8C, and passed through a 045-nm membrane and stored at 3-8C
Antibodies
Polysera
Antibody type and class
IgG and IgA
Assay System
Radioimmunoassay
Cross-reactivity
N/A
Proposed epitopes
N\A
Proposed Utility
These antibodies exert their protective effect by initiating complement-mediated activities including opsonization and bacterial lysis
