Mouse Anti-Lewis Y Recombinant Antibody (clone MSL5) (CAT#: FAMAB-0207CQ)

Figure 2 ELISA

A, Functional ELISA of mAbs MSL5 IgM (circles) and HI 8A lgG3 (squares) with Ley-BSA (white symbols) and Lex-BSA Ag (black symbols). The addition of 1 mM of LeY Ag to either H18A and MSL5 (10-40 pg/ml each) resulted in more than 95% reduction of the binding signal to Ley-BSA in each case. Binding of the Abs to milk powder, which was used as the blocking agent, was not detectable (inhibition and milk-powder data not shown). B, Functional ELISA of the tetrameric anti-LeY mini-antibody MSLS-p53his (circles) and dimeric anti-LeY mini-antibody MSL5-dHLXhis (squares) with LeYBSA (white symbols) and Lex-BSA Ag (black symbols). The addition of 1 mM of LeY Ag to both mini-antibody constructs (100-330 p.g/nil each) resulted in more than 95% reduction of the binding signal to Ley-BSA in each case. Binding of the Abs to milk powder, which was used as the blocking agent, was not detectable (inhibition and milkpowder data not shown). Bound Abs were detected with either IgM, IgG, or FLAG-specific alkaline phosphatase conjugates.

Rheinnecker, M., Hardt, C., Ilag, L. L., Kufer, P., Gruber, R., Hoess, A., ... & Pack, P. (1996). Multivalent antibody fragments with high functional affinity for a tumor-associated carbohydrate antigen. The Journal of Immunology, 157(7), 2989-2997.

Figure 3 Analysis of Ley-BSA-binding behavior of the MSL5-scFv (A), dimeric mini-antibody MSL5-dHLXhis (B), tetrameric mini-antibody MSL5-p53his (C), and MSL5-lgM (D) with surface plasmon resonance on a BlAcore biosensor instrument.

Shown are overlay plots of each construct at various concentrations (MSL5-scFv, 0.37-3.75 μM; dimeric mini-antibody MSL5-dHLXhis, 0.1 2-2.4 μM; tetrameric miniantibody MSL5-p53his, 0.08-1.07 μM; and IgM, 0.3-14 nM).

Rheinnecker, M., Hardt, C., Ilag, L. L., Kufer, P., Gruber, R., Hoess, A., ... & Pack, P. (1996). Multivalent antibody fragments with high functional affinity for a tumor-associated carbohydrate antigen. The Journal of Immunology, 157(7), 2989-2997.

Figure 4 Elution of MSL5-lgM (14 nM) from Ley-BSA using soluble LeY Ag.

The injection of 1 mM of LeY Ag is indicated by the arrow

Rheinnecker, M., Hardt, C., Ilag, L. L., Kufer, P., Gruber, R., Hoess, A., ... & Pack, P. (1996). Multivalent antibody fragments with high functional affinity for a tumor-associated carbohydrate antigen. The Journal of Immunology, 157(7), 2989-2997.

Figure 5 Overlay plot of individual dissociation curves of the MSL5-scFv fragment (3.5 μM) (A), dimeric mini-antibody MSL5-dHLXhis (2.4 μM) (B), tetrameric mini-antibody MSL5-p53his (1.07 μM) (C), and the MSL5-lgM (14 nM) (D).

Rheinnecker, M., Hardt, C., Ilag, L. L., Kufer, P., Gruber, R., Hoess, A., ... & Pack, P. (1996). Multivalent antibody fragments with high functional affinity for a tumor-associated carbohydrate antigen. The Journal of Immunology, 157(7), 2989-2997.