Defucosylated Anti-Epidermal Growth Factor Receptor Monoclonal Antibody (134-mG_2a-f) Exerts Antitumor Activities in Mouse Xenograft Models of Canine Osteosarcoma

Antitumor Activity of an anti-EGFR/HER2 Bispecific Antibody in a Mouse Xenograft Model of Canine Osteosarcoma

“…The H77scFv cDNA was further fused at the 3' end of the light chain cDNA of E134B. The cDNA of the E134B heavy chain and E134B light chain-H77scFv were transduced into BINDS-09 (FUT8 knockout ExpiCHO-S) cells using the ExpiCHO Expression System (Thermo Fisher Scientific, Inc.) to produce the defucosylated form [25][26][27][28][29][30][31][32]. A defucosylated and bispecific antibody, E134Bf-H77scFv, was purified using Ab-Capcher (ProteNova Co., Ltd.).…”

“…Canine mononuclear cells (MNCs) obtained from Yamaguchi University were resuspended in DMEM (Nacalai Tesque, Inc.) with 10% FBS and were used as effector cells. Target D-17 cells were labeled with 10 μg/mL Calcein AM (Thermo Fisher Scientific, Inc.) [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42]. The target cells (2 × 10 4 cells) were plated in 96-well plates and mixed with effector canine MNCs (effector/target cells ratio, 50), 100 μg/mL of control normal dog IgG, E134Bf, H77Bf, and E134Bf-H77scFv.…”

“…Target D-17 cells were treated with 10 μg/mL calcein AM [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42]. The target cells (2 × 10 4 cells) were mixed with rabbit complement (final dilution 1:10; Low-Tox-M Rabbit Complement; Cedarlane Laboratories) and 100 μg/mL of control normal dog IgG, E134Bf, H77Bf, and E134Bf-H77scFv.…”

“…Furthermore, on days 8, 15, and 22, canine MNCs (5 × 10 5 cells) were injected into the surrounding tumors. The tumor volume was measured on days 8, 10, 15, 17, 22, and 25, and determined, as described previously [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43].…”

An Anti-EpCAM Monoclonal Antibody (EpMab-37-mG2a-f) Exerts Antitumor Activity against Breast Cancer in Mouse Xenograft Model

“…VL cDNA of EpMab-37 and CL cDNA of mouse kappa light chain were also subcloned into the pCAG-Neo vector (Wako). To generate the defucosylated EpMab-37-mG2a, the vector for the EpMab-37-mG2a was transduced into BINDS-09 (FUT8 knockout ExpiCHO-S) cells using the ExpiCHO Expression System (Thermo) [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42]. Defucosytaled EpMab-37-mG2a (EpMab-37-mG2a-f) was purified using Ab-Capcher (ProteNova Co., Ltd., Kanagawa, Japan).…”