Anti-S. agalactiae FbsA protein Recombinant Antibody (5H2) (MRO-128MZ) (CAT#: MRO-128MZ)

Figure 1 Inhibition of Fng binding to 6pGST by the 5H2mAb.

Goat anti-GST was immobilized on the wells of microtiterplates, followed by incubation with 6pGST (100 nM). Fng (500nM) was mixed with 1 µg/ml of mAb 5H2, mAb 10H1 or mouseIgG1 (isotype control) before being added to the wells. Afterwashing, Fng binding was detected using rabbit anti-Fngantibodies followed by alkaline phosphatase-conjugated goatanti-rabbit IgG.

Immunogenic Properties of Streptococcus agalactiae FbsA Fragments

Figure 2 Inhibition of GBS attachment to surface-coatedFng by the 5H2 mAb.

Cells of S. agalactiae 6313 (5x107) werepreincubated with the indicated amounts of mAbs 5H2 or 10H1,transferred to Fng-coated wells (1 µg/well) and the mixtureswere incubated for 2 hours. After extensive washes, 1 µg rabbitanti-GBS IgG was added to the wells, followed by a 90 minincubation. Adherent bacteria were detected by peroxidase-conjugated goat anti-rabbit IgG and the plates were developedwith o-phenylenediamine. All the data are expressed aspercentages of control adherence, where bacteria attachmentin the absence of antibody was set to 100% (equivalent to 0%inhibition). The bars show standard deviations of triplicatesamples. This experiment was performed three times withsimilar results.

Immunogenic Properties of Streptococcus agalactiae FbsA Fragments

Figure 3 Effect of mAbs, directed against the repeat region of FbsA, on the interaction of S agalactiae 6313 or FbsA with human Fbg.

Concentration-dependent binding of the anti-FbsA mAbs 5H2 (▪) and 2B1 (⋄) to FbsA. Microtiter wells were coated overnight with 500 ng FbsA in 50 mM sodium carbonate, pH 9.5. The wells were treated for 1 hour at 22°C with 200 μL PBS containing 2% bovine serum albumin (BSA) and washed 5 times with PBS with 0.1% vol/vol Tween 20 (PBST). The plates were incubated with increasing amounts of mAbs 5H2 or 2B1 and the binding of each antibody was detected by incubating the wells with a 1:1000 dilution of rabbit antimouse peroxidase-conjugated polyclonal antibody. After washing, binding was quantitated using the substrate o-phenylenediamine dihydrochloride and measuring the absorbance at 490 nm.

FbsA, a fibrinogen-binding protein from Streptococcus agalactiae, mediates platelet aggregation