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Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering

Background: Native cluster of differentiation (CD) 19 targeting antibodies are poorly effective in triggering antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), which are crucial effector functions of therapeutic antibodies in cancer immunotherapy. Both...

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Autores principales: Roßkopf, Sophia, Eichholz, Klara Marie, Winterberg, Dorothee, Diemer, Katarina Julia, Lutz, Sebastian, Münnich, Ira Alexandra, Klausz, Katja, Rösner, Thies, Valerius, Thomas, Schewe, Denis Martin, Humpe, Andreas, Gramatzki, Martin, Peipp, Matthias, Kellner, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7709100/
https://www.ncbi.nlm.nih.gov/pubmed/33212776
http://dx.doi.org/10.3390/antib9040063
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author Roßkopf, Sophia
Eichholz, Klara Marie
Winterberg, Dorothee
Diemer, Katarina Julia
Lutz, Sebastian
Münnich, Ira Alexandra
Klausz, Katja
Rösner, Thies
Valerius, Thomas
Schewe, Denis Martin
Humpe, Andreas
Gramatzki, Martin
Peipp, Matthias
Kellner, Christian
author_facet Roßkopf, Sophia
Eichholz, Klara Marie
Winterberg, Dorothee
Diemer, Katarina Julia
Lutz, Sebastian
Münnich, Ira Alexandra
Klausz, Katja
Rösner, Thies
Valerius, Thomas
Schewe, Denis Martin
Humpe, Andreas
Gramatzki, Martin
Peipp, Matthias
Kellner, Christian
author_sort Roßkopf, Sophia
collection PubMed
description Background: Native cluster of differentiation (CD) 19 targeting antibodies are poorly effective in triggering antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), which are crucial effector functions of therapeutic antibodies in cancer immunotherapy. Both functions can be enhanced by engineering the antibody’s Fc region by altering the amino acid sequence (Fc protein-engineering) or the Fc-linked glycan (Fc glyco-engineering). We hypothesized that combining Fc glyco-engineering with Fc protein-engineering will rescue ADCC and CDC in CD19 antibodies. Results: Four versions of a CD19 antibody based on tafasitamab’s V-regions were generated: a native IgG1, an Fc protein-engineered version with amino acid exchanges S267E/H268F/S324T/G236A/I332E (EFTAE modification) to enhance CDC, and afucosylated, Fc glyco-engineered versions of both to promote ADCC. Irrespective of fucosylation, antibodies carrying the EFTAE modification had enhanced C1q binding and were superior in inducing CDC. In contrast, afucosylated versions exerted an enhanced affinity to Fcγ receptor IIIA and had increased ADCC activity. Of note, the double-engineered antibody harboring the EFTAE modification and lacking fucose triggered both CDC and ADCC more efficiently. Conclusions: Fc glyco-engineering and protein-engineering could be combined to enhance ADCC and CDC in CD19 antibodies and may allow the generation of antibodies with higher therapeutic efficacy by promoting two key functions simultaneously.
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spelling pubmed-77091002020-12-03 Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering Roßkopf, Sophia Eichholz, Klara Marie Winterberg, Dorothee Diemer, Katarina Julia Lutz, Sebastian Münnich, Ira Alexandra Klausz, Katja Rösner, Thies Valerius, Thomas Schewe, Denis Martin Humpe, Andreas Gramatzki, Martin Peipp, Matthias Kellner, Christian Antibodies (Basel) Article Background: Native cluster of differentiation (CD) 19 targeting antibodies are poorly effective in triggering antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), which are crucial effector functions of therapeutic antibodies in cancer immunotherapy. Both functions can be enhanced by engineering the antibody’s Fc region by altering the amino acid sequence (Fc protein-engineering) or the Fc-linked glycan (Fc glyco-engineering). We hypothesized that combining Fc glyco-engineering with Fc protein-engineering will rescue ADCC and CDC in CD19 antibodies. Results: Four versions of a CD19 antibody based on tafasitamab’s V-regions were generated: a native IgG1, an Fc protein-engineered version with amino acid exchanges S267E/H268F/S324T/G236A/I332E (EFTAE modification) to enhance CDC, and afucosylated, Fc glyco-engineered versions of both to promote ADCC. Irrespective of fucosylation, antibodies carrying the EFTAE modification had enhanced C1q binding and were superior in inducing CDC. In contrast, afucosylated versions exerted an enhanced affinity to Fcγ receptor IIIA and had increased ADCC activity. Of note, the double-engineered antibody harboring the EFTAE modification and lacking fucose triggered both CDC and ADCC more efficiently. Conclusions: Fc glyco-engineering and protein-engineering could be combined to enhance ADCC and CDC in CD19 antibodies and may allow the generation of antibodies with higher therapeutic efficacy by promoting two key functions simultaneously. MDPI 2020-11-17 /pmc/articles/PMC7709100/ /pubmed/33212776 http://dx.doi.org/10.3390/antib9040063 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Roßkopf, Sophia
Eichholz, Klara Marie
Winterberg, Dorothee
Diemer, Katarina Julia
Lutz, Sebastian
Münnich, Ira Alexandra
Klausz, Katja
Rösner, Thies
Valerius, Thomas
Schewe, Denis Martin
Humpe, Andreas
Gramatzki, Martin
Peipp, Matthias
Kellner, Christian
Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title_full Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title_fullStr Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title_full_unstemmed Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title_short Enhancing CDC and ADCC of CD19 Antibodies by Combining Fc Protein-Engineering with Fc Glyco-Engineering
title_sort enhancing cdc and adcc of cd19 antibodies by combining fc protein-engineering with fc glyco-engineering
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7709100/
https://www.ncbi.nlm.nih.gov/pubmed/33212776
http://dx.doi.org/10.3390/antib9040063
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