PolysacDB Glucan 1003
From DrugPedia: A Wikipedia for Drug discovery
Carbohydrate Name
Glucan
Carbohydrate Class
Miscellaneous
Source Microbe
Mycobacterium tuberculosis
Basic Structure
Glucans are a heterogeneous group of glucose polymers, consisting of a backbone of β(1-->3)-linked β-D-glucopyranosyl units with β(1-->6)-linked side chains of varying distribution and length
Proposed functions
Activates leukocytes, stimulating their phagocytic, cytotoxic, and antimicrobial activities, including the production of reactive oxygen and nitrogen intermediates
Antigenic Nature used to produce antibodies
Glycoconjugates
Carrier Name
Pseudomonas aeruginosa exoprotein A (rEPA)
Conjugation Method
The polysaccharide was activated with CDAP. A 60-ml volume of CDAP (100 mg/ml of acetonitrile) was added to a solution of polysaccharide (2 ml, 10 mg of polysaccharide per ml of PFS) at room temperature. The pH was maintained at 5.8 to 6.0 for 30 s, and 60 ml of 0.2 M TEA was added to a pH of 7.0. The reaction was carried out for 2 min, and an equal volume of 0.8 M adipic acid dihydrazide [ADH] in 0.5 M NaHCO3 was added. This reaction was carried out for 2 h, and the pH was maintained at 8.0 to 8.5 with 0.1 N NaOH. The reaction mixture was dialyzed against PFS and passed through a column (3 by 46 cm) of P-10 in PFW. The void volume fractions were pooled, freeze-dried. ADH-derivatized polysaccharide (10 mg) was dissolved in PFS (2 ml). An equal weight of protein was added, and the pH was maintained at 5.1 to 5.5 with 0.1 M HCl. The reaction mixture was put on ice, EDAC was added to a final concentration of 0.05 M, and the pH was maintained at 5.1 to 5.5 for 4 h in 0.1 M HCl. The reaction mixtures were dialyzed against 0.2 M NaCl for 2 days with three changes of outer fluid and were passed through a column (1.5 by 90 cm) of Sepharose CL-6B in 0.2 M NaCl. The void volume fractions were stored at 3 to 88C
Antibodies
Mab 24c5
Antibody type and class
IgG
Assay System
ELISA, Immunofluorescence
Cross-reactivity
This antibody cross-reacted with M. kansasii, BCG Pasteur, M. smegmatis, M. phlei, M. fortuitum, and M. avium
Proposed epitopes
N\A
Proposed Utility
The antibody was used to study M. tuberculosis Glucan expression in vitro and in vivo
