Defucosylated Anti-Epidermal Growth Factor Receptor Monoclonal Antibody 134-mG_2a-f Exerts Antitumor Activities in Mouse Xenograft Models of Dog Epidermal Growth Factor Receptor-Overexpressed Cells

Defucosylated Mouse–Dog Chimeric Anti-EGFR Antibody Exerts Antitumor Activities in Mouse Xenograft Models of Canine Tumors

“…CHO-K1, a canine osteosarcoma cell line (D-17 [ 17 ]), and a canine fibroblast cell line (A-72 [ 18 ]) were obtained from the American Type Culture Collection (Manassas, VA, USA). CHO/dEGFR was established in our previous study [ 16 ]. CHO-K1 and CHO/dEGFR were cultured in RPMI medium (Nacalai Tesque, Inc., Kyoto, Japan), D-17 in Minimum Essential Medium (Nacalai Tesque, Inc., Kyoto, Japan), and A-72 in Dulbecco’s Modified Eagle Medium (DMEM; Nacalai Tesque, Inc., Kyoto, Japan).…”

“…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 ]. The resulting mAb, E134Bf, was purified with Protein G-Sepharose (GE Healthcare Biosciences, Pittsburgh, PA, USA) [ 16 ].…”

“…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 ]. The resulting mAb, E134Bf, was purified with Protein G-Sepharose (GE Healthcare Biosciences, Pittsburgh, PA, USA) [ 16 ]. Dog IgG was purchased from Jackson ImmunoResearch Inc. (West Grove, PA, USA).…”

“…The mouse IgG 2a version of EMab-134 (134-mG 2a ) demonstrates antitumor activities in mouse xenograft models of hEGFR-expressing oral squamous cell carcinoma [ 15 ]. In addition, we produced the defucosylated version of 134-mG 2a (134-mG 2a -f) to augment antibody-dependent cellular cytotoxicity (ADCC) [ 16 ]. The 134-mG 2a -f exhibits ADCC and complement-dependent cytotoxicity (CDC) in dog EGFR (dEGFR)-overexpressed CHO-K1 (CHO/dEGFR) cells and antitumor activities in mouse xenograft models of CHO/dEGFR cells [ 16 ].…”

“…In addition, we produced the defucosylated version of 134-mG 2a (134-mG 2a -f) to augment antibody-dependent cellular cytotoxicity (ADCC) [ 16 ]. The 134-mG 2a -f exhibits ADCC and complement-dependent cytotoxicity (CDC) in dog EGFR (dEGFR)-overexpressed CHO-K1 (CHO/dEGFR) cells and antitumor activities in mouse xenograft models of CHO/dEGFR cells [ 16 ]. In this study, we investigated the antitumor activities of a defucosylated mouse–dog anti-EGFR mAb (E134Bf) against D-17 and A-72 xenografts, both of which endogenously express dEGFR.…”