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Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody

Monoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or mor...

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Autores principales: Sebastião, Maria João, Hoffman, Michael, Escandell, José, Tousi, Fatemeh, Zhang, Jin, Figueroa, Bruno, DeMaria, Christine, Gomes-Alves, Patrícia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669571/
https://www.ncbi.nlm.nih.gov/pubmed/38001891
http://dx.doi.org/10.3390/biomedicines11112890
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author Sebastião, Maria João
Hoffman, Michael
Escandell, José
Tousi, Fatemeh
Zhang, Jin
Figueroa, Bruno
DeMaria, Christine
Gomes-Alves, Patrícia
author_facet Sebastião, Maria João
Hoffman, Michael
Escandell, José
Tousi, Fatemeh
Zhang, Jin
Figueroa, Bruno
DeMaria, Christine
Gomes-Alves, Patrícia
author_sort Sebastião, Maria João
collection PubMed
description Monoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or more different polypeptide chains, MsAb production can lead to incorrect chain assembly and co-production of mispaired species with impaired biological activity. Moreover, mispairing carries significant challenges for downstream purification, decreasing yields and increasing the cost of bioprocess development. In this study, quantitative transcriptomics and proteomics analyses were employed to investigate which signaling pathways correlated with low and high mispairing clone signatures. Gene and protein expression profiles of Chinese hamster ovary (CHO) clones producing an tsAb were analyzed in the exponential growth and stationary (tsAb production) phase of fed-batch culture. Functional analysis revealed activated endoplasmic reticulum stress in high mispairing clones in both culture phases, while low mispairing clones exhibited expression profiles indicative of activated protein translation, as well as higher endocytosis and target protein degradation, suggesting the clearance of unfolded proteins through ubiquitin-mediated mechanisms. In addition, through transcriptomic profiling, we identified a group of genes that have the potential to be used as a biomarker panel tool for identifying high mispairing levels in the early stages of bioprocess development.
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spelling pubmed-106695712023-10-25 Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody Sebastião, Maria João Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia Biomedicines Article Monoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or more different polypeptide chains, MsAb production can lead to incorrect chain assembly and co-production of mispaired species with impaired biological activity. Moreover, mispairing carries significant challenges for downstream purification, decreasing yields and increasing the cost of bioprocess development. In this study, quantitative transcriptomics and proteomics analyses were employed to investigate which signaling pathways correlated with low and high mispairing clone signatures. Gene and protein expression profiles of Chinese hamster ovary (CHO) clones producing an tsAb were analyzed in the exponential growth and stationary (tsAb production) phase of fed-batch culture. Functional analysis revealed activated endoplasmic reticulum stress in high mispairing clones in both culture phases, while low mispairing clones exhibited expression profiles indicative of activated protein translation, as well as higher endocytosis and target protein degradation, suggesting the clearance of unfolded proteins through ubiquitin-mediated mechanisms. In addition, through transcriptomic profiling, we identified a group of genes that have the potential to be used as a biomarker panel tool for identifying high mispairing levels in the early stages of bioprocess development. MDPI 2023-10-25 /pmc/articles/PMC10669571/ /pubmed/38001891 http://dx.doi.org/10.3390/biomedicines11112890 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sebastião, Maria João
Hoffman, Michael
Escandell, José
Tousi, Fatemeh
Zhang, Jin
Figueroa, Bruno
DeMaria, Christine
Gomes-Alves, Patrícia
Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title_full Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title_fullStr Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title_full_unstemmed Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title_short Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
title_sort identification of mispairing omic signatures in chinese hamster ovary (cho) cells producing a tri-specific antibody
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10669571/
https://www.ncbi.nlm.nih.gov/pubmed/38001891
http://dx.doi.org/10.3390/biomedicines11112890
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