PolysacDB: A comprehensive database of microbial polysaccharide antigens and their antibodies

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Citation: Aithal A, Sharma A, Joshi S, Raghava GPS, Varshney GC (2012) PolysacDB: A Database of Microbial Polysaccharide Antigens and Their Antibodies. PLoS ONE 7(4): e34613. doi:10.1371/journal.pone.0034613
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PolysacDB ID1030
Carbohydrate NameO-deacetylated Glucurunoxylomannan   (Drugpedia)
Carbohydrate ClassCapsular polysaccharide
Microbe NameCryptococcus neoformans Serotype C   (NCBI Taxonomy)   (Drugpedia)
Basic StructureThe GXM is a high-molecular-weight linear (1-->3)-α-D-mannan substituted with single (1-->2)-β-xylosyl and (1-->2)-β-glucuronosyl residues. 0 acetylation occurs on the C-6 of about half of the mannose residues. The molar ratio of xylose-mannose-glucuronic acid is 2:3:1 for serotype A
BCSDB Structure73
Proposed FunctionThe capsular glucuronoxylomannan (GXM) of C. neoformans is anantiphagocytic and tolerogenic polysaccharide and potentiates virulence
Antigenic NatureGlycoconjugates
Carrier NameBovine serum albumin
Conjugate MethodGXM of C. neoformans serotype A 9759 was activated with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride) and was coupled to BSA-AH. GXM (100 mg) was suspended in acetone, and 75 ul of pyridine was added. Tresyl chloride (50 ,ul) was added and stirred for 10 min. The activated GXM was washed with ethanol containing 5 mM HCI, washed with absolute ethanol, and dried in vacuo. Activated GXM (46 mg) was dissolved in 1.5 ml of 0.2 M NaH2PO4 buffer (pH 7.5) (coupling buffer), and BSA-AH was added (25 mg in 0.5 ml of the same buffer). The reaction was stirred overnight at 24°C. The precipitate was removed by centrifugation, and the supernatant was applied to a column (90 by 2.5 cm) of Sepharose CL-6B equilibrated with 0.05 M Tris hydrochloride-0.1 M NaCl buffer (pH 7.6). The flow rate was 35 ml/h. Fractions containing carbohydrate and protein appeared in the void volume. They were pooled, dialyzed, and lyophilized (yield, 33 mg). The conjugate was then rechromatographed (yield, 17 mg)
AntibodiesMab CD6
Antibody Type ClassIgG1
Assay SystemIndirect immunofluorescence
Cross ReactivityThis antibody cross-reacted with all 4 serotypes- A, B, C and D.
Proposed EpitopeGlucuronic acid was proposed to be the important constituent of the epitope. However the epitope also probably contained xylose and was influencedto a minor extent by the presence of O-acetyl groups
IEDB EpitopeN/A
Proposed UtilityThe ability to produce a MAb against an epitope shared by many serotypes may have value for the detection of cryptococcal antigens in body fluids. This antibody also has potent opsonic activity
Curator IDAA + AS
Date of Curation09-01-2010
ReferencesPMC259921

Bioinformatics Centre, Institute of Microbial Technology, Sec - 39A, Chandigarh, India - 160036