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Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy

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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.
Keywords
IGG ANTIBODIES, CHAIN ASSOCIATION, GROWTH-FACTOR, TUMOR-CELLS, T-CELLS, FRAGMENTS, CANCER, CHROMATOGRAPHY, BISPECIFIC ANTIBODIES, SINGLE-DOMAIN ANTIBODY

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Chicago
Godar, Marie, Virginia Morello, Ava Sadi, Anna Hultberg, Natalie De Jonge, Cristina Basilico, Valérie Hanssens, et al. 2016. “Dual Anti-idiotypic Purification of a Novel, Native-format Biparatopic anti-MET Antibody with Improved in Vitro and in Vivo Efficacy.” Scientific Reports 6.
APA
Godar, M., Morello, V., Sadi, A., Hultberg, A., De Jonge, N., Basilico, C., Hanssens, V., et al. (2016). Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy. SCIENTIFIC REPORTS, 6.
Vancouver
1.
Godar M, Morello V, Sadi A, Hultberg A, De Jonge N, Basilico C, et al. Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy. SCIENTIFIC REPORTS. 2016;6.
MLA
Godar, Marie, Virginia Morello, Ava Sadi, et al. “Dual Anti-idiotypic Purification of a Novel, Native-format Biparatopic anti-MET Antibody with Improved in Vitro and in Vivo Efficacy.” SCIENTIFIC REPORTS 6 (2016): n. pag. Print.
@article{8132438,
  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.},
  articleno    = {31621},
  author       = {Godar, Marie and Morello, Virginia and Sadi, Ava and Hultberg, Anna and De Jonge, Natalie and Basilico, Cristina and Hanssens, Val{\'e}rie and Saunders, Michael and Lambrecht, Bart and El Khattabi, Mohamed and de Haard, Hans and Michieli, Paolo and Blanchetot, Christophe},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  keyword      = {IGG ANTIBODIES,CHAIN ASSOCIATION,GROWTH-FACTOR,TUMOR-CELLS,T-CELLS,FRAGMENTS,CANCER,CHROMATOGRAPHY,BISPECIFIC ANTIBODIES,SINGLE-DOMAIN ANTIBODY},
  language     = {eng},
  pages        = {12},
  title        = {Dual anti-idiotypic purification of a novel, native-format biparatopic anti-MET antibody with improved in vitro and in vivo efficacy},
  url          = {http://dx.doi.org/10.1038/srep31621},
  volume       = {6},
  year         = {2016},
}

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