Generation of anti-EGFR mAb 134-mG2a

Anti-EGFR mAb EMab-134 (mouse IgG₁, kappa) was developed as previously described (14 (link)). To generate 134-mG_2a, V_H cDNA of EMab-134 and C_H mouse IgG_2a were subcloned into pCAG-Ble vector, and V_L and C_L cDNAs of EMab-134 were subcloned into pCAG-Neo vector (FUJIFILM Wako Pure Chemical Corporation), respectively. Vectors were transfected into ExpiCHO-S cells using the ExpiCHO Expression System (Thermo Fisher Scientific, Inc.). The resulting mAb, 134-mG_2a, was purified with Protein G-Sepharose (GE Healthcare Bio-Sciences). Mouse IgG (cat. no. I8765), IgG₁ (cat. no. M7894), and IgG_2a (cat. no. M7769) were purchased from Sigma-Aldrich; Merck KGaA.