Temperature Upshifts in Mammalian Cell Culture: A Suitable Strategy for Biosimilar Monoclonal Antibodies?

Lukas Marschall; Chitti Babu Gottimukkala; Biswajit Kayal; Veerabhadra Madurai Veeraraghavan; Samir Kumar Mandal; Suman Bandyopadhyay; Christoph Herwig

doi:10.3390/bioengineering10101149

Temperature Upshifts in Mammalian Cell Culture: A Suitable Strategy for Biosimilar Monoclonal Antibodies?

Bioengineering (Basel). 2023 Sep 30;10(10):1149. doi: 10.3390/bioengineering10101149.

Authors

Lukas Marschall^{1

2}, Chitti Babu Gottimukkala³, Biswajit Kayal³, Veerabhadra Madurai Veeraraghavan³, Samir Kumar Mandal³, Suman Bandyopadhyay³, Christoph Herwig^{1

2}

Affiliations

¹ TU Wien, Faculty of Technical Chemistry, Research Unit Biochemical Engineering, Gumpendorferstrasse 1a, 1060 Vienna, Austria.
² Körber Pharma Austria GmbH, Mariahilfer Straße 88A/1/9, 1070 Vienna, Austria.
³ Dr. Reddy's Laboratories Ltd., Biologics, Survey No. 47, Bachupally, Hyderabad 500090, India.

Abstract

Temperature downshifts are the gold standard when setting up control strategies for mammalian cell culture processes. These shifts are performed to prolong production phases and attain heightened levels of productivity. For the development of biosimilars, however, the bottleneck is in achieving a prespecified product quality. In a late-stage development project, we investigated the impact of temperature shifts and other process parameters with the aim of optimizing the glycosylation profile of a monoclonal antibody (mAb). We applied a design of experiments approach on a 3 L scale. The optimal glycosylation profile was achieved when performing a temperature upshift from 35.8 °C to 37 °C. Total afucosylated glycan (TAF) decreased by 1.2%, and galactosylated glycan species (GAL) increased by up to 4.5%. The optimized control strategy was then successfully taken to the manufacturing scale (1000 L). By testing two sets of set points at the manufacturing scale, we demonstrated that the statistical models predicting TAF and GAL trained with small-scale data are representative of the manufacturing scale. We hope this study encourages researchers to widen the screening ranges in process development and investigate whether temperature upshifts are also beneficial for other mAbs.

Keywords: biosimilar; mammalian cell culture; scale-up; temperature downshift.

Grants and funding

Open Access Funding by TU Wien. The authors acknowledge TU Wien Bibliothek for financial support for editing/proofreading. The authors acknowledge TU Wien Bibliothek for financial support through its Open Access Funding Program. The project, which forms the basis of this work, was a commercial collaboration funded by Dr Reddy’s Laboratories Ltd. in collaboration with Körber Pharma Austria GmbH.