Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells

Abstract One major challenge observed for the expression of therapeutic bispecific antibodies (BisAbs) is high product aggregates. Aggregates increase the risk of immune responses in patients and therefore must be removed at the expense of purification yields. BisAbs contain engineered disulfide bon...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Pritam Sinharoy, Aaron H. Aziz, Natalia I. Majewska, Sanjeev Ahuja, Michael W. Handlogten
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8cab4892c86c4270b94b3c96cdbbf671
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8cab4892c86c4270b94b3c96cdbbf671
record_format	dspace
spelling	oai:doaj.org-article:8cab4892c86c4270b94b3c96cdbbf6712021-12-02T18:37:06ZPerfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells10.1038/s41598-020-73573-42045-2322https://doaj.org/article/8cab4892c86c4270b94b3c96cdbbf6712020-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-73573-4https://doaj.org/toc/2045-2322Abstract One major challenge observed for the expression of therapeutic bispecific antibodies (BisAbs) is high product aggregates. Aggregates increase the risk of immune responses in patients and therefore must be removed at the expense of purification yields. BisAbs contain engineered disulfide bonds, which have been demonstrated to form product aggregates, if mispaired. However, the underlying intracellular mechanisms leading to product aggregate formation remain unknown. We demonstrate that impaired glutathione regulation underlies BisAb aggregation formation in a CHO cell process. Aggregate formation was evaluated for the same clonal CHO cell line producing a BisAb using fed-batch and perfusion processes. The perfusion process produced significantly lower BisAb aggregates compared to the fed-batch process. Perfusion bioreactors attenuated mitochondrial dysfunction and ER stress resulting in a favorable intracellular redox environment as indicated by improved reduced to oxidized glutathione ratio. Conversely, mitochondrial dysfunction-induced glutathione oxidation and ER stress disrupted the intracellular redox homeostasis, leading to product aggregation in the fed-batch process. Combined, our results demonstrate that mitochondrial dysfunction and ER stress impaired glutathione regulation leading to higher product aggregates in the fed-batch process. This is the first study to utilize perfusion bioreactors as a tool to demonstrate the intracellular mechanisms underlying product aggregation formation.Pritam SinharoyAaron H. AzizNatalia I. MajewskaSanjeev AhujaMichael W. HandlogtenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Pritam Sinharoy Aaron H. Aziz Natalia I. Majewska Sanjeev Ahuja Michael W. Handlogten Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
description	Abstract One major challenge observed for the expression of therapeutic bispecific antibodies (BisAbs) is high product aggregates. Aggregates increase the risk of immune responses in patients and therefore must be removed at the expense of purification yields. BisAbs contain engineered disulfide bonds, which have been demonstrated to form product aggregates, if mispaired. However, the underlying intracellular mechanisms leading to product aggregate formation remain unknown. We demonstrate that impaired glutathione regulation underlies BisAb aggregation formation in a CHO cell process. Aggregate formation was evaluated for the same clonal CHO cell line producing a BisAb using fed-batch and perfusion processes. The perfusion process produced significantly lower BisAb aggregates compared to the fed-batch process. Perfusion bioreactors attenuated mitochondrial dysfunction and ER stress resulting in a favorable intracellular redox environment as indicated by improved reduced to oxidized glutathione ratio. Conversely, mitochondrial dysfunction-induced glutathione oxidation and ER stress disrupted the intracellular redox homeostasis, leading to product aggregation in the fed-batch process. Combined, our results demonstrate that mitochondrial dysfunction and ER stress impaired glutathione regulation leading to higher product aggregates in the fed-batch process. This is the first study to utilize perfusion bioreactors as a tool to demonstrate the intracellular mechanisms underlying product aggregation formation.
format	article
author	Pritam Sinharoy Aaron H. Aziz Natalia I. Majewska Sanjeev Ahuja Michael W. Handlogten
author_facet	Pritam Sinharoy Aaron H. Aziz Natalia I. Majewska Sanjeev Ahuja Michael W. Handlogten
author_sort	Pritam Sinharoy
title	Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
title_short	Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
title_full	Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
title_fullStr	Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
title_full_unstemmed	Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells
title_sort	perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and er stress in cho cells
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/8cab4892c86c4270b94b3c96cdbbf671
work_keys_str_mv	AT pritamsinharoy perfusionreducesbispecificantibodyaggregationviamitigatingmitochondrialdysfunctioninducedglutathioneoxidationanderstressinchocells AT aaronhaziz perfusionreducesbispecificantibodyaggregationviamitigatingmitochondrialdysfunctioninducedglutathioneoxidationanderstressinchocells AT nataliaimajewska perfusionreducesbispecificantibodyaggregationviamitigatingmitochondrialdysfunctioninducedglutathioneoxidationanderstressinchocells AT sanjeevahuja perfusionreducesbispecificantibodyaggregationviamitigatingmitochondrialdysfunctioninducedglutathioneoxidationanderstressinchocells AT michaelwhandlogten perfusionreducesbispecificantibodyaggregationviamitigatingmitochondrialdysfunctioninducedglutathioneoxidationanderstressinchocells
_version_	1718377810364465152

Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells

Ejemplares similares