In situ cell differentiation monitoring of Catharanthus roseus suspension culture processes by NIR spectroscopy.

Daniel Arturo Zavala-Ortiz,Annie Marc,Dulce María Barradas-Dermitz,Patricia Margaret Hayward-Jones,María Guadalupe Aguilar-Uscanga,Emmanuel Guedon,Bruno Ebel

doi:10.1007/s00449-019-02255-x

Abstract

Plant suspension culture is attracting interest as a promising platform to produce biological medicines due to the absence of virus, prions or DNA related to mammals during the production process. However, the heterogenic plant cell proliferation nature is particularly challenging for establishing industrial processes based on innovative approaches currently used, particularly in the animal cell culture industry. In this context, while Process Analytical Technology (PAT) tools have been used to monitor classical parameters such as biomass dry weight, its use in cells heterogeneity has received limited attention. Therefore, the feasibility of in situ monitoring of cell differentiation in plant cell suspensions employing NIR spectroscopy and chemometrics was investigated. Off-line measurements of cell heterogeneity in term of cell differentiation and in-line NIR spectra captured in 3 L bioreactor cultures were employed to generate calibration models. Then models were tested to estimate the population distribution of parenchyma, collenchyma and sclerenchyma cells during Catharanthus roseus suspension cultures. Results have proven in situ NIR spectroscopy as a capable PAT tool to monitor differentiated cells accurately and in real-time. These results are the starting point to follow-up PAT systems so that plant cell culture heterogeneity may be better understood and controlled in biopharmaceutical plant cell cultures.

Highlights

Plant cell suspension cultures have attracted interest in biological medicine production due mainly to safety aspects [1]
This study lays the foundation for future studies to expand the capabilities of Process Analytical Technology (PAT) tools for in situ and real-time monitoring of cell heterogeneity in biopharmaceutical production processes based on plant cell suspension cultures
Determination of latent variable (LV) number was based on the goodness of estimation (Q2Y), which is the fraction of the captured variation of the cell concentration estimated by the model

Summary

Introduction

Plant cell suspension cultures have attracted interest in biological medicine production due mainly to safety aspects [1]. This study lays the foundation for future studies to expand the capabilities of PAT tools for in situ and real-time monitoring of cell heterogeneity in biopharmaceutical production processes based on plant cell suspension cultures. In this context, NIR spectroscopy could be a monitoring platform for real-time monitoring of the state of cells first, providing meaningful information of cell physiological state using calibration models, and second, performing retro-control strategies, leading to a more efficient design and control of plant suspension culture processes

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Conclusion