Anti-EGFR monoclonal antibody 134-mg_2a exerts antitumor effects in mouse xenograft models of oral squamous cell carcinoma

The epidermal growth factor receptor (EGFR), a transmembrane receptor and member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases, is a critical mediator of cell growth and differentiation. EGFR forms homo- or heterodimers with other HER family members to activate downstream signaling cascades in a number of cancer cells. In a previous study, the authors established an anti-EGFR monoclonal antibody (mAb), EMab-134, by immunizing mice with the ectodomain of human EGFR. EMab-134 binds specifically to endogenous EGFR and can be used to detect receptor on oral cancer cell lines by flow cytometry and western blot analysis; this antibody is also effective for the immunohistochemical evaluation of oral cancer tissues. In the present study, the subclass of EMab-134 was converted from IgG₁ to IgG_2a (134-mG_2a) to facilitate antibody-dependent cellular cytotoxicity (Adcc) and complement-dependent cytotoxicity (cdc). The dissociation constants (K_ds) of EMab-134 and 134-mG_2a against EGFR-expressing cHO-K1 (cHO/EGFR) cells were determined by flow cytometry to be 3.2x10^-9 M and 2.1x10^-9 M, respectively; these results indicate that 134-mG_2a has a higher binding affinity than EMab-134. The 134-mG_2a antibody was more sensitive than EMab-134 with respect to antigen detection in oral cancer cells in both western blot analysis and immunohistochemistry applications. Analysis in vitro revealed that 134-mG_2a contributed to high levels of Adcc and cdc in experiments targeting cHO/EGFR, HSc-2, and SAS cells. Moreover, the in vivo administration of 134-mG_2a significantly inhibited the development of cHO/EGFR, HSc-2, and SAS mouse xenografts in comparison to the results observed in response to EMab-134. Taken together, the findings of the present study demonstrate that the newly-formulated 134-mG_2a is useful for detecting EGFR by flow cytometry, western blot analysis and immunohistochemistry. Furthermore, the in vivo results suggested that it may also be useful as part of a therapeutic regimen for patients with EGFR-expressing oral cancer.