Human Anti-Tetanus toxoid Recombinant Antibody (clone F13) (CAT#: VS-0623-WK14)

Figure 1 The reactivity of the Fab fragment with the tetanus toxoid in immuno blot analysis.

The purified F13 was electrophoresed on a 12% SDS-PAGE. The resolved proteins were transferred to a polyvinyl difluoride membrane following the manufacturer's instructions. The residual sites were blocked using 2% skimmed milk powder and probed with anti-human IgG (Fab-specific) HRPO 1:5000 (Sigma, St. Louis, USA). The blot was developed using 0.05% 3,3-diaminobenzidine tetrahydrochloride (DAB) (Sigma, USA) and 0.03% hydrogen peroxide in PBS.

Figure 2 Analysis of antigen-binding affinity of F13 Fab by Indirect ELISA to evaluate the activity of Fab toward tetanus toxoid.

The F13 Fab showed signifi cant binding activity at high concentration (2 μg/100 μl) with the OD of ~1.0 and gradually reduced to ~0.1 with the decrease in concentration.

Figure 3 Analysis of F13 Fab activity toward tetanus toxoid in sandwich ELISA.

2F8 Mouse mAb was coated as a capture antibody, and titration of tetanus toxoid antigen against a constant dilution of F13 Fab revealed a concentration-dependent reduction of the binding signal in the wells. The E. coli lysate used
as a negative control showed negligible binding activity with tetanus toxoid antigen in the ELISA plate.

Figure 4 Competitive ELISA using the F13 Fab and the mouse monoclonal antibody.

Constant OD values (around ~1.0) in the dilution of both antibodies indicated that the F13 Fab and mouse mAb were binding to different antigenic sites on tetanus toxoid.

Figure 5 Specificity of F13 Fab toward various Clostridium species toxins.

Figure 6 Neutralizing efficiency of F13 Fab toward tetanus toxoid by flocculation assay.

Neutralization expressed in the limit of flocculation.