Real time monitoring of bioreactor mAb IgG3 cell culture process dynamics via Fourier transform infrared spectroscopy: Implications for enabling cell culture process analytical technologyŽ 

Abstract

Compared to small molecule process analytical technology (PAT) applications, biotechnology product PAT applications have certain unique challenges and opportunities. Understanding process dynamics of bioreactor cell culture process is essential to establish an appropriate process control strategy for biotechnology product PAT applications. Inline spectroscopic techniques for real time monitoring of bioreactor cell culture process have the distinct potential to develop PAT approaches in manufacturing biotechnology drug products. However, the use of inline Fourier transform infrared (FTIR) spectroscopic techniques for bioreactor cell culture process monitoring has not been reported. In this work, real time inline FTIR Spectroscopy was applied to a lab scale bioreactor mAb IgG3 cell culture fluid biomolecular dynamic model. The technical feasibility of using FTIR Spectroscopy for real time tracking and monitoring four key cell culture metabolites (including glucose, glutamine, lactate, and ammonia) and protein yield at increasing levels of complexity (simple binary system, fully formulated media, actual bioreactor cell culture process) was evaluated via a stepwise approach. The FTIR fingerprints of the key metabolites were identified. The multivariate partial least squares (PLS) calibration models were established to correlate the process FTIR spectra with the concentrations of key metabolites and protein yield of in-process samples, either individually for each metabolite and protein or globally for all four metabolites simultaneously. Applying the 2^nd derivative pre-processing algorithm to the FTIR spectra helps to reduce the number of PLS latent variables needed significantly and thus simplify the interpretation of the PLS models. The validated PLS models show promise in predicting the concentration profiles of glucose, glutamine, lactate, and ammonia and protein yield over the course of the bioreactor cell culture process. Therefore, this work demonstrated the technical feasibility of real time monitoring of the bioreactor cell culture process via FTIR spectroscopy. Its implications for enabling cell culture PAT were discussed.

Graphical abstract

Keywords

process analytical technology (PAT) / Fourier-transform infrared (FTIR) spectroscopy / partial least squares (PLS) regression / mouse IgG3 / bioreactor cell culture process / real time process monitoring

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Acknowledgements

This work was financially supported by FDA Center for Drug Evaluation and Research (CDER) Regulatory Science and Review (RSR) funding RSR-12-42, RSR-13-32, FDA CDER Office of Testing and Research (OTR) Funding Wu-12-PAT, and FDA CDER OBP’s PAT Critical Path project 1500. The unmatched technical support from Mettler-Toledo AutoChem team including but not limited to Vaso Vlachos, Paul Scholl, Simon Rea, and Jack Sue in the aspects of ReactIR probe autoclaving and ReactIR system optimization is greatly acknowledged. H. Wu wishes to thank Dr. Jennifer Maguire, Dr. Rapti Madurawe, Dr. Christine Moore, Dr. Vincent Vilker (retired), Dr. Patrick Faustino, Dr. Lucinda Buhse, and Dr. Lawrence Yu at the Office of Process and Facilities (OPF), Office of Pharmaceutical Quality (OPQ), Center for Drug Evaluation and Research (CDER), FDA for their management supports during the preparation and finalization of this manuscript. H. Wu is grateful to Prof. Jingkang Wang, Dr. Yingjin Yuan, Dr. Zhenhua Li at Tianjin University for the kind invitation, to Dr. Xiaowen Zhu for prompt communication and hard work, and to Yanni Li and Luli Cheng for their editorial assistance.

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2015 Higher Education Press and Springer-Verlag Berlin Heidelberg