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Identification of Anti-EGFR and Anti-ErbB3 Dual Variable Domains Immunoglobulin (DVD-Ig) Proteins with Unique Activities.

Gu J, Yang J, Chang Q, Liu Z, Ghayur T, Gu J - PLoS ONE (2015)

Bottom Line: Interestingly, in the presence of β-Heregulin (HRG), the DVD-Ig proteins show synergies with respect to inhibiting cell proliferation.The DVD-Ig proteins downregulate EGFR protein expression in the presence of HRG, which may be due to receptor internalization.Furthermore, the DVD-Ig proteins remarkably disrupt β-Heregulin binding to FaDu cells.

View Article: PubMed Central - PubMed

Affiliation: AbbVie Bioresearch Center, R&D, Worcester, Massachusetts, 01605, United States of America.

ABSTRACT
Epidermal growth factor receptor (EGFR) and receptor tyrosine-protein kinase 3 (ErbB3) are two well-established targets in cancer therapy. There is significant crosstalk among these two receptors and others. To block signaling from both EGFR and ErbB3, we generated anti-EGFR and anti-ErbB3 DVD-Ig proteins. Two DVD-Ig proteins maintained the functions of the combination of the two parental antibodies. The DVD-Ig proteins inhibit cell signaling and proliferation in A431 and FaDu cells while half DVD-Ig proteins lost proliferation inhibition function. Interestingly, in the presence of β-Heregulin (HRG), the DVD-Ig proteins show synergies with respect to inhibiting cell proliferation. The DVD-Ig proteins downregulate EGFR protein expression in the presence of HRG, which may be due to receptor internalization. Furthermore, the DVD-Ig proteins remarkably disrupt β-Heregulin binding to FaDu cells.

No MeSH data available.


Related in: MedlinePlus

DVD-Ig proteins inhibit HRG binding to FaDu cells.Serial dilutions of HRG were incubated with 100 nM anti-EGFR and anti-ErbB3 mAbs alone, anti-EGFR and anti-ErbB3 mAbs in combination, the bsAb, or anti-EGFR/ErbB3 DVD-Ig proteins. HRG binding affinity to FaDu cells was measured via FACS. MFI: median fluorescence intensity. The error bars indicate standard deviation from the mean.
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pone.0124135.g006: DVD-Ig proteins inhibit HRG binding to FaDu cells.Serial dilutions of HRG were incubated with 100 nM anti-EGFR and anti-ErbB3 mAbs alone, anti-EGFR and anti-ErbB3 mAbs in combination, the bsAb, or anti-EGFR/ErbB3 DVD-Ig proteins. HRG binding affinity to FaDu cells was measured via FACS. MFI: median fluorescence intensity. The error bars indicate standard deviation from the mean.

Mentions: To test this hypothesis, we analyzed the binding affinity of Alexa488-labeled HRG to FaDu cells in the presence of the anti-EGFR and anti-ErbB3 mAbs alone, the anti-EGFR and anti-ErbB3 mAbs in combination, and the anti-EGFR/ErbB3 DVD-Ig proteins. The results showed that both DVD1 and DVD2 proteins could block HRG binding to FaDu cells more efficiently (Fig 6). For example, when incubated with 10nM HRG, 100nM DVD1 and DVD2 could block >90% of HRG binding to the cells, while 100nM anti-ErbB3 mAb, the bsAb, or the combination could only block ~75% of HRG binding to the cells (Fig 6).


Identification of Anti-EGFR and Anti-ErbB3 Dual Variable Domains Immunoglobulin (DVD-Ig) Proteins with Unique Activities.

Gu J, Yang J, Chang Q, Liu Z, Ghayur T, Gu J - PLoS ONE (2015)

DVD-Ig proteins inhibit HRG binding to FaDu cells.Serial dilutions of HRG were incubated with 100 nM anti-EGFR and anti-ErbB3 mAbs alone, anti-EGFR and anti-ErbB3 mAbs in combination, the bsAb, or anti-EGFR/ErbB3 DVD-Ig proteins. HRG binding affinity to FaDu cells was measured via FACS. MFI: median fluorescence intensity. The error bars indicate standard deviation from the mean.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440733&req=5

pone.0124135.g006: DVD-Ig proteins inhibit HRG binding to FaDu cells.Serial dilutions of HRG were incubated with 100 nM anti-EGFR and anti-ErbB3 mAbs alone, anti-EGFR and anti-ErbB3 mAbs in combination, the bsAb, or anti-EGFR/ErbB3 DVD-Ig proteins. HRG binding affinity to FaDu cells was measured via FACS. MFI: median fluorescence intensity. The error bars indicate standard deviation from the mean.
Mentions: To test this hypothesis, we analyzed the binding affinity of Alexa488-labeled HRG to FaDu cells in the presence of the anti-EGFR and anti-ErbB3 mAbs alone, the anti-EGFR and anti-ErbB3 mAbs in combination, and the anti-EGFR/ErbB3 DVD-Ig proteins. The results showed that both DVD1 and DVD2 proteins could block HRG binding to FaDu cells more efficiently (Fig 6). For example, when incubated with 10nM HRG, 100nM DVD1 and DVD2 could block >90% of HRG binding to the cells, while 100nM anti-ErbB3 mAb, the bsAb, or the combination could only block ~75% of HRG binding to the cells (Fig 6).

Bottom Line: Interestingly, in the presence of β-Heregulin (HRG), the DVD-Ig proteins show synergies with respect to inhibiting cell proliferation.The DVD-Ig proteins downregulate EGFR protein expression in the presence of HRG, which may be due to receptor internalization.Furthermore, the DVD-Ig proteins remarkably disrupt β-Heregulin binding to FaDu cells.

View Article: PubMed Central - PubMed

Affiliation: AbbVie Bioresearch Center, R&D, Worcester, Massachusetts, 01605, United States of America.

ABSTRACT
Epidermal growth factor receptor (EGFR) and receptor tyrosine-protein kinase 3 (ErbB3) are two well-established targets in cancer therapy. There is significant crosstalk among these two receptors and others. To block signaling from both EGFR and ErbB3, we generated anti-EGFR and anti-ErbB3 DVD-Ig proteins. Two DVD-Ig proteins maintained the functions of the combination of the two parental antibodies. The DVD-Ig proteins inhibit cell signaling and proliferation in A431 and FaDu cells while half DVD-Ig proteins lost proliferation inhibition function. Interestingly, in the presence of β-Heregulin (HRG), the DVD-Ig proteins show synergies with respect to inhibiting cell proliferation. The DVD-Ig proteins downregulate EGFR protein expression in the presence of HRG, which may be due to receptor internalization. Furthermore, the DVD-Ig proteins remarkably disrupt β-Heregulin binding to FaDu cells.

No MeSH data available.


Related in: MedlinePlus