Identification of anti-ErbB2 dual variable domain immunoglobulin (DVD-Ig™) proteins with unique activities.

Jinming Gu,Jijie Gu,Xiaoqing Lu,Tariq Ghayur,Jieyi Wang,Qing Chang,Jinsong Yang,Mingjiu Chen,Geetha P Bansal

doi:10.1371/journal.pone.0097292

Abstract

Inhibiting ErbB2 signaling with monoclonal antibodies (mAbs) or small molecules is an established therapeutic strategy in oncology. We have developed anti-ErbB2 Dual Variable Domain Immunoglobulin (DVD-Ig) proteins that capture the function of a combination of two anti-ErbB2 antibodies. In addition, some of the anti-ErbB2 DVD-Ig proteins gain the new functions of enhancing ErbB2 signaling and cell proliferation in N87 cells. We further found that two DVD-Ig proteins, DVD687 and DVD688, have two distinct mechanisms of actions in Calu-3 and N87 cells. DVD687 enhances cell cycle progression while DVD688 induces apoptosis in N87 cells. Using a half DVD687, we found that avidity may play a key role in the agonist activity of DVD687 in N87 cells.

Highlights

ErbB2 is one of the four members of the ErbB family of receptor tyrosine kinases (RTKs)
Generation of Anti-ErbB2 Dual Variable Domain Immunoglobulin (DVD-Ig) Proteins To test whether we could capture the synergistic effect of two anti-ErbB2 antibodies via the DVD-Ig platform, we used the variable domains of two different anti-ErbB2 antibodies to generate eight DVD-Ig proteins differing in the orientation of the two variable domains and linkers used (Table 1)
The results show that all eight DVD-Ig proteins have similar binding affinity to ErbB2-extra cellular domain (ECD) as compared to single antibodies (Table 2)

Summary

Introduction

ErbB2 is one of the four members of the ErbB family of receptor tyrosine kinases (RTKs). The ErbB family members have multiple ligands, including epidermal growth factor (EGF), Heregulin, Betacellulin, and TGFa [10,11,12]. Upon ligand binding, they form homodimers and/or heterodimers, which induce receptor internalization and/ or intracellular signaling [11,13]. There is a significant amount of crosstalk among ErbB family members and other cell receptor tyrosine kinases, such as cMet and IGF1R, in cancer progression and drug resistance [14,15,16,17,18,19]

Methods

Results

Conclusion