Recombinant Mouse Anti-F. tularensis LPS Antibody (Ab52) (CAT#: PABC-178)

Figure 1 Mouse Anti-F. tularensis LPS Antibody (Ab52) in ELISA.

All four anti-LPS IgG2a MAbs are specific for virulent type A strain by ELISA. ELISA plates were coated with heat-killed (HK) SchuS4 (A) or TG1, as described in Materials and Methods. IgG2a MAbs Ab53 (anti-GroEL), CO17-1A (anti-human EpCam), and GTX40330 (mouse monoclonal IgG2a to Gram negative endotoxin) were used as specificity and isotype controls.

Roche, M. I., Lu, Z., Hui, J. H., & Sharon, J. (2011). Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide. Hybridoma, 30(1), 19-28.

Figure 2 Mouse Anti-F. tularensis LPS Antibody (Ab52) in ELISA.

All four anti-LPS IgG2a MAbs are specific for OAg by ELISA. ELISA plates were coated with heat-killed (HK) LVS, Ft LPS, HK WbtI, or Ft OAg, as described in Materials and Methods. IgG2a MAbs Ab53 (anti-GroEL), CO17-1A (anti-human EpCam), and mIgG2a (anti-non-human PBLs) were used as specificity and isotype controls.

Roche, M. I., Lu, Z., Hui, J. H., & Sharon, J. (2011). Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide. Hybridoma, 30(1), 19-28.

Figure 3 Mouse Anti-F. tularensis LPS Antibody (Ab52) in IP.

FB11 cannot immunoprecipitate OAg despite its higher bacterial agglutination titer compared to the other three MAbs. Immunoprecipitation by double immunodiffusion in agar. The contrast and brightness of the photograph were globally increased to allow better visualization of the thin precipitin lines formed with Ab3 and Ab54, which were clearly visible with the naked eye.

Roche, M. I., Lu, Z., Hui, J. H., & Sharon, J. (2011). Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide. Hybridoma, 30(1), 19-28.

Figure 4 Mouse Anti-F. tularensis LPS Antibody (Ab52) in SPR.

The affinity of FB11, but not of Ab52, can be determined using immobilized MAb and soluble Ft OAg. Schematic of the monovalent binding of immobilized FB11 to Ft OAg (top) and of the multivalent binding of immobilized Ab52 to Ft OAg (bottom).

Roche, M. I., Lu, Z., Hui, J. H., & Sharon, J. (2011). Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide. Hybridoma, 30(1), 19-28.

Figure 5 Mouse Anti-F. tularensis LPS Antibody (Ab52) in SPR.

The avidity of FB11 is higher than those of Ab52 and Ab54. Biacore analysis of the binding of soluble FB11, Ab52, and Ab54 to immobilized Ft LPS. Ab52, captured on anti-mouse IgG-coated chips, was used to capture Ft LPS, which was then probed with FB11, Ab52, or Ab54. MAb concentration ranges used in the assays were 0 − 10 nM for FB11, 0 − 20 nM for Ab52, and 0 − 100 nM for Ab54.

Roche, M. I., Lu, Z., Hui, J. H., & Sharon, J. (2011). Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide. Hybridoma, 30(1), 19-28.