title: Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy.
abstract: Bispecific antibodies are of great interest due to their ability to simultaneously bind and engage different antigens or epitopes. Nevertheless, it remains a challenge to assemble, produce and/or purify them. Here we present an innovative dual anti-idiotypic purification process, which provides pure bispecific antibodies with native immunoglobulin format. Using this approach, a biparatopic IgG1 antibody targeting two distinct, HGF-competing, non-overlapping epitopes on the extracellular region of the MET receptor, was purified with camelid single-domain antibody fragments that bind specifically to the correct heavy chain/light chain pairings of each arm. The purity and functionality of the anti-MET biparatopic antibody was then confirmed by mass spectrometry and binding experiments, demonstrating its ability to simultaneously target the two epitopes recognized by the parental monoclonal antibodies. The improved MET-inhibitory activity of the biparatopic antibody compared to the parental monoclonal antibodies, was finally corroborated in cell-based assays and more importantly in a tumor xenograft mouse model. In conclusion, this approach is fast and specific, broadly applicable and results in the isolation of a pure, novel and native-format anti-MET biparatopic antibody that shows superior biological activity over the parental monospecific antibodies both in vitro and in vivo.
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.|VIB Inflammation Research Center 9052 Zwijnaarde, Belgium.|Department of Internal Medicine, Ghent University, 9000 Ghent, Belgium.
authors.forename: Marie
authors.initials: M
authors.lastname: Godar
authors.affiliation: Department of Oncology, University of Torino Medical School, 10060 Candiolo, Turin, Italy.|Candiolo Cancer Institute, FPO-IRCCS, 10060 Candiolo, Turin, Italy.
authors.forename: Virginia
authors.initials: V
authors.lastname: Morello
authors.affiliation: QVQ BV, Yalelaan 1 Androclus building, 3584 CL Utrecht, The Netherlands.
authors.forename: Ava
authors.initials: A
authors.lastname: Sadi
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Anna
authors.initials: A
authors.lastname: Hultberg
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Natalie
authors.initials: N
authors.lastname: De Jonge
authors.affiliation: Candiolo Cancer Institute, FPO-IRCCS, 10060 Candiolo, Turin, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Basilico
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Valérie
authors.initials: V
authors.lastname: Hanssens
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Michael
authors.initials: M
authors.lastname: Saunders
authors.affiliation: VIB Inflammation Research Center 9052 Zwijnaarde, Belgium.|Department of Internal Medicine, Ghent University, 9000 Ghent, Belgium.|Department of Pulmonary Medicine, ErasmusMC, 3015 GE Rotterdam, The Netherlands.
authors.forename: Bart N
authors.initials: BN
authors.lastname: Lambrecht
authors.affiliation: QVQ BV, Yalelaan 1 Androclus building, 3584 CL Utrecht, The Netherlands.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El Khattabi
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Hans
authors.initials: H
authors.lastname: de Haard
authors.affiliation: Department of Oncology, University of Torino Medical School, 10060 Candiolo, Turin, Italy.|Candiolo Cancer Institute, FPO-IRCCS, 10060 Candiolo, Turin, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Michieli
authors.affiliation: argenx BVBA, Industriepark Zwijnaarde 7, Building C, 9052 Zwijnaarde, Belgium.
authors.forename: Christophe
authors.initials: C
authors.lastname: Blanchetot
mesh_terms: D000072283:A549 Cells; D000818:Animals; D018033:Antibodies, Bispecific / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D000074322:Antineoplastic Agents, Immunological / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D016513:Mice, SCID; D009374:Neoplasms, Experimental / Q000188:drug therapy* / Q000276:immunology; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
chemical_list: D018033:Antibodies, Bispecific; D000074322:Antineoplastic Agents, Immunological; D007074:Immunoglobulin G; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met