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An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells

Complex therapeutic antibody formats, such as bispecifics (bsAbs) or cytokine fusions, may provide new treatment options in diverse disease areas. However, the manufacturing yield of these complex antibody formats in Chinese Hamster Ovary (CHO) cells is lower than monoclonal antibodies due to challe...

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Autores principales: Bauer, Niels, Oswald, Benedikt, Eiche, Maximilian, Schiller, Lisa, Langguth, Emma, Schantz, Christian, Osterlehner, Andrea, Shen, Amy, Misaghi, Shahram, Stingele, Julian, Ausländer, Simon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700384/
https://www.ncbi.nlm.nih.gov/pubmed/36452067
http://dx.doi.org/10.1093/synbio/ysac026
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author Bauer, Niels
Oswald, Benedikt
Eiche, Maximilian
Schiller, Lisa
Langguth, Emma
Schantz, Christian
Osterlehner, Andrea
Shen, Amy
Misaghi, Shahram
Stingele, Julian
Ausländer, Simon
author_facet Bauer, Niels
Oswald, Benedikt
Eiche, Maximilian
Schiller, Lisa
Langguth, Emma
Schantz, Christian
Osterlehner, Andrea
Shen, Amy
Misaghi, Shahram
Stingele, Julian
Ausländer, Simon
author_sort Bauer, Niels
collection PubMed
description Complex therapeutic antibody formats, such as bispecifics (bsAbs) or cytokine fusions, may provide new treatment options in diverse disease areas. However, the manufacturing yield of these complex antibody formats in Chinese Hamster Ovary (CHO) cells is lower than monoclonal antibodies due to challenges in expression levels and potential formation of side products. To overcome these limitations, we performed a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-based knockout (KO) arrayed screening of 187 target genes in two CHO clones expressing two different complex antibody formats in a production-mimicking set-up. Our findings revealed that Myc depletion drastically increased product expression (>40%) by enhancing cell-specific productivity. The Myc-depleted cells displayed decreased cell densities together with substantially higher product titers in industrially-relevant bioprocesses using ambr15 and ambr250 bioreactors. Similar effects were observed across multiple different clones, each expressing a distinct complex antibody format. Our findings reinforce the mutually exclusive relationship between growth and production phenotypes and provide a targeted cell engineering approach to impact productivity without impairing product quality. We anticipate that CRISPR/Cas9-based CHO host cell engineering will transform our ability to increase manufacturing yield of high-value complex biotherapeutics.
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spelling pubmed-97003842022-11-29 An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells Bauer, Niels Oswald, Benedikt Eiche, Maximilian Schiller, Lisa Langguth, Emma Schantz, Christian Osterlehner, Andrea Shen, Amy Misaghi, Shahram Stingele, Julian Ausländer, Simon Synth Biol (Oxf) Research Article Complex therapeutic antibody formats, such as bispecifics (bsAbs) or cytokine fusions, may provide new treatment options in diverse disease areas. However, the manufacturing yield of these complex antibody formats in Chinese Hamster Ovary (CHO) cells is lower than monoclonal antibodies due to challenges in expression levels and potential formation of side products. To overcome these limitations, we performed a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-based knockout (KO) arrayed screening of 187 target genes in two CHO clones expressing two different complex antibody formats in a production-mimicking set-up. Our findings revealed that Myc depletion drastically increased product expression (>40%) by enhancing cell-specific productivity. The Myc-depleted cells displayed decreased cell densities together with substantially higher product titers in industrially-relevant bioprocesses using ambr15 and ambr250 bioreactors. Similar effects were observed across multiple different clones, each expressing a distinct complex antibody format. Our findings reinforce the mutually exclusive relationship between growth and production phenotypes and provide a targeted cell engineering approach to impact productivity without impairing product quality. We anticipate that CRISPR/Cas9-based CHO host cell engineering will transform our ability to increase manufacturing yield of high-value complex biotherapeutics. Oxford University Press 2022-11-03 /pmc/articles/PMC9700384/ /pubmed/36452067 http://dx.doi.org/10.1093/synbio/ysac026 Text en © The Author(s) 2022. Published by Oxford University Press. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (https://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Research Article
Bauer, Niels
Oswald, Benedikt
Eiche, Maximilian
Schiller, Lisa
Langguth, Emma
Schantz, Christian
Osterlehner, Andrea
Shen, Amy
Misaghi, Shahram
Stingele, Julian
Ausländer, Simon
An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title_full An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title_fullStr An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title_full_unstemmed An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title_short An arrayed CRISPR screen reveals Myc depletion to increase productivity of difficult-to-express complex antibodies in CHO cells
title_sort arrayed crispr screen reveals myc depletion to increase productivity of difficult-to-express complex antibodies in cho cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700384/
https://www.ncbi.nlm.nih.gov/pubmed/36452067
http://dx.doi.org/10.1093/synbio/ysac026
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