Generating Anti-hEGFR Monoclonal Antibody E134B

The anti-hEGFR mAb, EMab-134, was developed as previously described [14 (link)]. To produce E134B, we subcloned the V_H cDNA of EMab-134 and C_H cDNA of Dog IgGB into the pCAG-Ble vector (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), along with the V_L cDNA of EMab-134 and C_L cDNA of dog kappa light chain into the pCAG-Neo vector (FUJIFILM Wako Pure Chemical Corporation). Two vectors of E134B were transfected into BINDS-09 cells (FUT8-deficient ExpiCHO-S cells) using the ExpiCHO Expression System (Thermo Fisher Scientific Inc., Waltham, MA, USA) [16 (link)]. The resulting mAb, E134Bf, was purified with Protein G-Sepharose (GE Healthcare Biosciences, Pittsburgh, PA, USA) [16 (link)]. Dog IgG was purchased from Jackson ImmunoResearch Inc. (West Grove, PA, USA).