Abstract
Animal cell culture processes have become the standard platform to produce therapeutic proteins such as recombinant monoclonal antibodies (mAb). Since the mAb quality could be subject to significant changes depending on manufacturing process conditions, real time monitoring and control systems are required to ensure mAb specifications mainly glycosylation and patient safety. Up to now, real time monitoring glycosylation of proteins has received scarce attention. In this article, the use of near infrared (NIR) to monitor mAb glycosylation has been reported for the first time. Whereas monitoring models are mainly constructed using linear partial least squares regressions (PLSR), evidences presented in this study indicate nonlinearity relationship between in situ captured spectra and compound concentrations, compromising the PLSR performances. A novel and simple approach was proposed to fit nonlinearity using the locally weighted regression (LWR). The LWR models were found to be more appropriate for handling information contained in spectra so that real time monitoring of cultures were accurately performed. Moreover, for the first time, the LWR calibration models allowed mAb glycosylation to be monitored, in a real time manner, by using in situ NIR spectroscopy. These results represent a further step toward developing active-control feedback of animal cell processes, particularly for ensuring properties of biologics.
Highlights
The production of biologicals, especially recombinant monoclonal antibodies, remains a challenge due to the structural complexity of these molecules and their sensitivity to changes in the manufacturing process
Models for total monoclonal antibodies (mAb) and nonglycosylated mAb (NG-mAb) concentrations were generated with the aim of monitoring the quality of mAbs produced during a process
The widely used partial least squares regressions (PLSR) method was incapable of relating spectra with compound concentrations, indicating that such a widely used regression method is not always appropriate for the monitoring of animal cell culture processes
Summary
The production of biologicals, especially recombinant monoclonal antibodies (mAb), remains a challenge due to the structural complexity of these molecules and their sensitivity to changes in the manufacturing process. That is why strict quality control systems are required to ensure mAb specifications and patient safety. As one of the main quality attributes, glycosylation pattern confers chemical and therapeutic properties to mAb (serum half-life, immunogenicity, antibody-dependent cellular cytotoxicity, and complementdependent cytotoxicity).[6,7,8] its control is essential to ensure efficacy of the product and safety for patients. A new challenging objective for PAT is to control mAb glycosylation as well as cell metabolism using online spectroscopy.[13,14,15,16] accurate monitoring models must be developed so that advanced active feedback control systems for controlling processes could become feasible
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