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Application of in situ near infra-red spectroscopy (NIRS) for monitoring biopharmaceuticals production by cell cultures

Abstract : The complexity of biopharmaceutical products implies that their approval is based on a specific process. Any further change, especially in the process, requires drug validation in terms of clinical effects and biosecurity. Because changes in the processes may be unavoidable, quality assurance by inspection at the end of the process (Quality by Testing-QbT) tends to be replaced by a new quality perspective called Quality by design (QbD) which builds drug Critical Quality Attributes (CQA) controlling key Critical Process Parameters (CPP) in real-time. However, QbD implementation has been limited by the complexity of cell culture processes and the need for multivariate methods that allow the use of complex signals from process analyzers as monitoring instruments. Consequently, the objective of this work has been to develop new methodological and experimental applications, based on in situ NIR spectroscopy, for real-time monitoring of biopharmaceutical-producing cell cultures using two production platforms: animal cells (CHO-250-9) producing monoclonal antibodies (mAb) and plant cells (Cantharanthus roseus) producing antineoplastic molecules (vincristine-VC and vinblastine-VB). First, a process capable of producing VC and VB was generated, cell differentiation was identified as CPP and the ability to monitor it by in situ NIR spectroscopy was firstly demonstrated using calibration models based on partial least squares regression (PLSR). Subsequently in CHO cell cultures, different regression techniques were evaluated to generate calibration models to monitor CPP and CQA. PLSR was inadequate because of the chemical and physical variability that CHO cell cultures present during the different phases of batch culture. Local Weighted Regression (LWR) was adequate to monitor classic CPP (concentration of glucose, lactate, and viable cells, amongst others) since it adequately handled the variability associated with the progression of cell culture. However, for the glycosylation profile (CQA), it was unable to properly handle the complex nonlinear relationships between NIR spectra and the concentration of various monoclonal antibody (mAb) glycoforms. This was overcome with the use of models based on support vector regressions (SVR), allowing the generation of models of different mAb glycoforms related to particular clinical effects. Globally, this work has contributed to the expansion of the capabilities of in situ NIR spectroscopy for the monitoring of classic CPP in a more precise way, new innovative CPP such as cell physiological state in plant suspension cultures, and CQA such as mAb glycosylation profiles linked to clinical characteristics in animal cell cultures.
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https://hal.univ-lorraine.fr/tel-02929681
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Daniel Zavala-Ortiz. Application of in situ near infra-red spectroscopy (NIRS) for monitoring biopharmaceuticals production by cell cultures. Chemical and Process Engineering. Université de Lorraine; Instituto tecnólogico de Veracruz (Mexique), 2020. English. ⟨NNT : 2020LORR0040⟩. ⟨tel-02929681⟩

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