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Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices

Generation of bispecific antibodies (bsAbs) requires a combination of compatible binders in formats that support desired functionalities. Here, we report that bsAb-matrices can be generated by Format Chain Exchange (FORCE), enabling screening of combinatorial binder/format spaces. Input molecules fo...

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Autores principales: Dengl, Stefan, Mayer, Klaus, Bormann, Felix, Duerr, Harald, Hoffmann, Eike, Nussbaum, Bianca, Tischler, Michael, Wagner, Martina, Kuglstatter, Andreas, Leibrock, Lea, Buldun, Can, Georges, Guy, Brinkmann, Ulrich
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7532213/
https://www.ncbi.nlm.nih.gov/pubmed/33009381
http://dx.doi.org/10.1038/s41467-020-18477-7
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author Dengl, Stefan
Mayer, Klaus
Bormann, Felix
Duerr, Harald
Hoffmann, Eike
Nussbaum, Bianca
Tischler, Michael
Wagner, Martina
Kuglstatter, Andreas
Leibrock, Lea
Buldun, Can
Georges, Guy
Brinkmann, Ulrich
author_facet Dengl, Stefan
Mayer, Klaus
Bormann, Felix
Duerr, Harald
Hoffmann, Eike
Nussbaum, Bianca
Tischler, Michael
Wagner, Martina
Kuglstatter, Andreas
Leibrock, Lea
Buldun, Can
Georges, Guy
Brinkmann, Ulrich
author_sort Dengl, Stefan
collection PubMed
description Generation of bispecific antibodies (bsAbs) requires a combination of compatible binders in formats that support desired functionalities. Here, we report that bsAb-matrices can be generated by Format Chain Exchange (FORCE), enabling screening of combinatorial binder/format spaces. Input molecules for generation of bi/multi-valent bsAbs are monospecific entities similar to knob-into-hole half-antibodies, yet with complementary CH3-interface-modulated and affinity-tagged dummy-chains. These contain mutations that lead to limited interface repulsions without compromising expression or biophysical properties of educts. Mild reduction of combinations of educts triggers spontaneous chain-exchange reactions driven by partially flawed CH3-educt interfaces resolving to perfect complementarity. This generates large bsAb matrices harboring different binders in multiple formats. Benign biophysical properties and good expression yields of educts, combined with simplicity of purification enables process automation. Examples that demonstrate the relevance of screening binder/format combinations are provided as a matrix of bsAbs that simultaneously bind Her1/Her2 and DR5 without encountering binder or format-inflicted interferences.
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spelling pubmed-75322132020-10-19 Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices Dengl, Stefan Mayer, Klaus Bormann, Felix Duerr, Harald Hoffmann, Eike Nussbaum, Bianca Tischler, Michael Wagner, Martina Kuglstatter, Andreas Leibrock, Lea Buldun, Can Georges, Guy Brinkmann, Ulrich Nat Commun Article Generation of bispecific antibodies (bsAbs) requires a combination of compatible binders in formats that support desired functionalities. Here, we report that bsAb-matrices can be generated by Format Chain Exchange (FORCE), enabling screening of combinatorial binder/format spaces. Input molecules for generation of bi/multi-valent bsAbs are monospecific entities similar to knob-into-hole half-antibodies, yet with complementary CH3-interface-modulated and affinity-tagged dummy-chains. These contain mutations that lead to limited interface repulsions without compromising expression or biophysical properties of educts. Mild reduction of combinations of educts triggers spontaneous chain-exchange reactions driven by partially flawed CH3-educt interfaces resolving to perfect complementarity. This generates large bsAb matrices harboring different binders in multiple formats. Benign biophysical properties and good expression yields of educts, combined with simplicity of purification enables process automation. Examples that demonstrate the relevance of screening binder/format combinations are provided as a matrix of bsAbs that simultaneously bind Her1/Her2 and DR5 without encountering binder or format-inflicted interferences. Nature Publishing Group UK 2020-10-02 /pmc/articles/PMC7532213/ /pubmed/33009381 http://dx.doi.org/10.1038/s41467-020-18477-7 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Dengl, Stefan
Mayer, Klaus
Bormann, Felix
Duerr, Harald
Hoffmann, Eike
Nussbaum, Bianca
Tischler, Michael
Wagner, Martina
Kuglstatter, Andreas
Leibrock, Lea
Buldun, Can
Georges, Guy
Brinkmann, Ulrich
Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title_full Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title_fullStr Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title_full_unstemmed Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title_short Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
title_sort format chain exchange (force) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7532213/
https://www.ncbi.nlm.nih.gov/pubmed/33009381
http://dx.doi.org/10.1038/s41467-020-18477-7
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