United Kingdom (UK)
Recombinant monoclonal antibody to 2.4[2.8]2.4-linked Kdo tetrasaccharide. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma S25-2.
Immunogen: This antibody was generated by immunizing a BALB/c mouse with KD03-GlcNAc-BSA.
Specificity: This antibody is specific for 2.4[2.8]2.4-linked Kdo tetrasaccharide. Which is a partial structure of chlamydial lipopolysaccharide (LPS).
Application Notes: An enzyme immunoassay was performed on LPS-BSA complexes and synthetic BSA-glycoconjugates using the mouse version of this antibody it showed reactivity to r595-207 LPS and to KD03-GlcNAc-BSA. Furthermore this antibody was used for immunofluorescence on L929 monolayers infected with C.trachomatis serotype L2 or C. psittaci 6BC at a dose resulting in >80% infected cells within 24 to 36 h, the antibody gave a positive result for both infections. Additionally western blots with LPS r595-207 and R595 as the antigen were performed using this antibody, it was postive for r595-207 and negative for R595 (Fu et al, 1992; pmid:1372290). An ELISA was performed on chlamydial LPS fragments using the mouse version of this antibody it bound most strongly to trisaccharides and tetrasaccharides (Evans et al, 2011; pmid:21543444). The binding affinity of the mouse version of this antibody was determined using SPR on α–Kdo-(2→8)-α–Kdo-(2→4)-α–Kdo, it bound with a Kd of 1,1μM (Müller-Loennies et al, 2000; pmid:10642603). SPR was performed on oligosaccharides using the mouse version of this antibody, for binding see Brooks et al, 2008. Furthermore ELISA was performed on neoglycoconjugates using this antibody, for reactivity see Brooks et al, 2008 (Brooks et al, 2008; pmid:18272175).
Antibody first published in:
Fu et al. A synthetic glycoconjugate representing the genus-specific epitope of chlamydial lipopolysaccharide exhibits the same specificity as its natural counterpart. Infect Immun. 1992 Apr;60(4):1314-21. PMID:1372290
Note on publication:
The tetrasaccharide 3-deoxy-a-D-manno-2-octulosonic acid (ac-KDO)(2--8)-ce-KDO(2->4)-ce-KDO(2-->6)- IGIcNAc, a partial structure of chlamydial lipopolysaccharide (LPS) representing a genus-specific epitope, was synthesized and covalently linked to bovine serum albumin, resulting in an artificial glycoconjugate antigen. Mice were immunized with the glycoconjugate to prepare chlamydia-specific monoclonal antibodies.