Human Anti-ERBB2 Recombinant Antibody (clone F5) (CAT#: HPAB-S0027-YC)

Figure 1 Receptor-mediated internalization of anti-ErbB2 antibodies into SkBr-3 cells.

Anti-ErbB2 scFv: monovalent C6.5 and F5, bivalent dbC6.5 and F5(scFv9 )2, in comparison to the anti-ErbB2 control antibodies, Herceptin/trastuzumab (Her) and 4D5Fab9 . Affinity constants determined on cells (Kd) and by BIAcore for ErbB2 are indicated below the graph. Cells were incubated with 125I-labeled antibodies at 37°C for 2 h. The amounts of cell surface-bound and internalized antibodies were determined by acid rinse technique, and antibody affinity constants (Kd values) for ErbB2 were determined by flow cytometry on SKOV3 cells.

Neve, R. M., Nielsen, U. B., Kirpotin, D. B., Poul, M. A., Marks, J. D., & Benz, C. C. (2001). Biological effects of anti-ErbB2 single chain antibodies selected for internalizing function. Biochemical and biophysical research communications, 280(1), 274-279.

Figure 2 (A) Cell cycle profiles of SkBr-3 cells. (B) Effect of monovalent and bivalent anti-ErbB2 scFv and control anti-ErbB2 antibody on SkBr-3 culture growth.

A. Neither the C6.5 nor F5 monovalent scFv had any significant affect on the SkBr-3 cell cycle profile relative to untreated control cells, while the bivalent forms of these scFv caused an apparent mitogenic cell cycle push reflected by an increase in the proportion of S-phase cells after 24 h of treatment. B. neither monovalent nor bivalent forms of C6.5 and F5 scFv result in any significant growth altering effects on ErbB2 overepressing cells.

Neve, R. M., Nielsen, U. B., Kirpotin, D. B., Poul, M. A., Marks, J. D., & Benz, C. C. (2001). Biological effects of anti-ErbB2 single chain antibodies selected for internalizing function. Biochemical and biophysical research communications, 280(1), 274-279.

Figure 3 Time-dependent responses in the MAPK and PI3K signaling pathways upon exposure of SkBr-3 cells to monovalent or bivalent scFv.

All antibodies had variable degrees of an early (30 min) and transient agonist-like effect on basal MAPK signaling, measured by increased levels of phospho-Erk1/Erk2 and phospho-JNK.

Neve, R. M., Nielsen, U. B., Kirpotin, D. B., Poul, M. A., Marks, J. D., & Benz, C. C. (2001). Biological effects of anti-ErbB2 single chain antibodies selected for internalizing function. Biochemical and biophysical research communications, 280(1), 274-279.

Figure 4 Effects of scFv pretreatment on SkBr-3 cell responses to the ErbB growth factor, NDF.

NDF-induced cell signaling in both SkBr-3 and MCF-7 cells occurs by MAPK (increased phospho-Erk1/Erk2 and phospho-JNK levels) and PI3K (increased phospho-PKB levels) transduction pathways.

Neve, R. M., Nielsen, U. B., Kirpotin, D. B., Poul, M. A., Marks, J. D., & Benz, C. C. (2001). Biological effects of anti-ErbB2 single chain antibodies selected for internalizing function. Biochemical and biophysical research communications, 280(1), 274-279.