Recombinant Mouse Anti-Staphylococcal enterotoxin B (SEB) Antibody (6D3) (CAT#: PABL-706)

Figure 1 ELISA shows the effect of binding using different site directed mutagenesis proteins.

Mutant proteins were coated in polystyrene plates at a concentration of 0.5 ug/ml. Further mAb 20B1 or 14G8 or 6D3 or 4C7 was added, detected by alkaline phosphatase (AP)-conjugated goat anti-mouse IgG1 and developed by PNPP tablets. The x -axis represents absorbance at 405 nm and y -axis represents the log of antibody concentration. Results identify different critical residues, which could interact with the individual SEB specific mAbs. For mAb 20B1 mutation of residue 135-R, 137-F, 186-Y, 235 & 236-T affected binding. The residues 135-R, 186-Y were required for the interaction with mAb 6D3. mAb 14G8 bind to residues 135-R, 137-F, 186-Y, 188-K, 231-E, 233-Y, and 235, 236-T, whereas mAb 4C7 interact with 135-R, 137-F, 186-Y, 188-K, and 235, 236-T.

Varshney, A. K. , Wang, X. , Cook, E. , Dutta, K. , & Fries, B. C. . (2011). Generation, characterization, and epitope mapping of neutralizing and protective monoclonal antibodies against staphylococcal enterotoxin b-induced lethal shock. Journal of Biological Chemistry, 286(11), 9737-9747.

Figure 2 Inhibition of T-cell proliferation and cytokine production by treatment with SEB specific mAb 20B1, 14G8, 6D3, and 4C7 individually or in different combinations.

A , SEB-induced T-cell proliferation was measured by ViaLight HS Cell Proliferation kit after 48 h ( A ) and 96 h ( B ) and inhibited in the presence of all three mAb except 4C7. IFN ( C ) and IL-2 ( D ) were measured by ELISA in the supernatant of SEB stimulated T-cells. Cytokines were significantly lower in the presence of mAbs relative to conditions with no specific antibody. The bars represent the S.D. derived from triplicate wells from same experiment.

Varshney, A. K. , Wang, X. , Cook, E. , Dutta, K. , & Fries, B. C. . (2011). Generation, characterization, and epitope mapping of neutralizing and protective monoclonal antibodies against staphylococcal enterotoxin b-induced lethal shock. Journal of Biological Chemistry, 286(11), 9737-9747.

Figure 3 Dot blot analysis of 14G8.

Dot blot analysis shows binding of 10-mer peptide with all three mAbs with SEB and mutant-1 and no binding with mutant-2.

Varshney, A. K. , Wang, X. , Cook, E. , Dutta, K. , & Fries, B. C. . (2011). Generation, characterization, and epitope mapping of neutralizing and protective monoclonal antibodies against staphylococcal enterotoxin b-induced lethal shock. Journal of Biological Chemistry, 286(11), 9737-9747.

Figure 1 Recombinant Mouse Anti-Staphylococcal enterotoxin B Antibody (PABL-706) in SDS-PAGE

Lane M: Marker
Lane 1: PABL-706 in non-reduced condition.
Lane 2: PABL-706 in reduced condition.

Figure 2 Recombinant Mouse Anti-Staphylococcal enterotoxin B Antibody (PABL-706) in HPLC

HPLC analysis of PABL-706 shows the purity is>95%.