A novel glycoengineered bispecific antibody format for targeted inhibition of epidermal growth factor receptor (EGFR) and insulin-like growth factor receptor type I (IGF-1R) demonstrating unique molecular properties.
In the present study, we have developed a novel one-arm single chain Fab heterodimeric bispecific IgG (OAscFab-IgG) antibody format targeting the insulin-like growth factor receptor type I (IGF-1R) and the epidermal growth factor receptor (EGFR) with one binding site for each target antigen. The bispecific antibody XGFR is based on the "knob-into-hole" technology for heavy chain heterodimerization with one heavy chain consisting of a single chain Fab to prevent wrong pairing of light chains. XGFR was produced with high expression yields and showed simultaneous binding to IGF-1R and EGFR with high affinity. Due to monovalent binding of XGFR to IGF-1R, IGF-1R internalization was strongly reduced compared with the bivalent parental antibody, leading to enhanced Fc-mediated cellular cytotoxicity. To further increase immune effector functions triggered by XGFR, the Fc portion of the bispecific antibody was glycoengineered, which resulted in strong antibody-dependent cell-mediated cytotoxicity activity. XGFR-mediated inhibition of IGF-1R and EGFR phosphorylation as well as A549 tumor cell proliferation was highly effective and was comparable with a combined treatment with EGFR (GA201) and IGF-1R (R1507) antibodies. XGFR also demonstrated potent anti-tumor efficacy in multiple mouse xenograft tumor models with a complete growth inhibition of AsPC1 human pancreatic tumors and improved survival of SCID beige mice carrying A549 human lung tumors compared with treatment with antibodies targeting either IGF-1R or EGFR. In summary, we have applied rational antibody engineering technology to develop a heterodimeric OAscFab-IgG bispecific antibody, which combines potent signaling inhibition with antibody-dependent cell-mediated cytotoxicity induction and results in superior molecular properties over two established tetravalent bispecific formats.
From Discovery Oncology and juergen_michael.schanzer@roche.com.
Juergen M
JM
Schanzer
From Discovery Oncology and.
Katharina
K
Wartha
From Discovery Oncology and.
Rebecca
R
Croasdale
Roche Glycart AG, CH-8952 Schlieren, Switzerland, and.
Samuel
S
Moser
Boehringer Ingelheim, A-1121 Vienna, Austria.
Klaus-Peter
KP
Künkele
From Discovery Oncology and.
Carola
C
Ries
From Discovery Oncology and.
Werner
W
Scheuer
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Harald
H
Duerr
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Sandra
S
Pompiati
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Jan
J
Pollman
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Jan
J
Stracke
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Wilma
W
Lau
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Stefan
S
Ries
Large Molecule Research, Pharmaceutical Research and Early Development, Roche Diagnostics GmbH, Nonnenwald 2, D-82372 Penzberg, Germany.
Ulrich
U
Brinkmann
Roche Glycart AG, CH-8952 Schlieren, Switzerland, and.
Christian
C
Klein
Roche Glycart AG, CH-8952 Schlieren, Switzerland, and.
Pablo
P
Umana
D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D066246:ErbB Receptors / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D010190:Pancreatic Neoplasms / Q000473:pathology; D015202:Protein Engineering; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D021381:Protein Transport / Q000187:drug effects; D017526:Receptor, IGF Type 1 / Q000276:immunology* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
D018033:Antibodies, Bispecific; D007074:Immunoglobulin G; D057127:Single-Chain Antibodies; D066246:ErbB Receptors; D017526:Receptor, IGF Type 1
Antibody Engineering; Epidermal Growth Factor Receptor (EGFR); Glycosylation; Insulin-like Growth Factor (IGF); Tumor Therapy