PolysacDB Capsular polysaccharide 1005

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Contents

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

Weblink

PubMed Central

External Links