BIOTHERAPEUTICS DEVELOPABILITY
AND RISK MITIGATION

PARTNERS

INSTITUTIONAL PARTNERS

MEDIA PARTNERS

Fabien Compeau

Design and development of chemically defined media supplemented with vegetable protein hydrolysates to enhance CHO cell growth and monoclonal antibody production

Abstract

In the realm of biopharmaceutical production, Chinese Hamster Ovary (CHO) cells have emerged as the main expression system for recombinant therapeutic proteins (RTPs), despite the availability of several alternative cell lines. CHO cells are among the most widely used worldwide for their unique advantages, e.g, the variety of post-translational modifications that closely mimic those found on human cells (Stach et al. 2019); their high degree of biosafety due to their low risk of propagation of human viruses infection (Lalonde and Durocher, 2017); their adaptability from adherent to suspension cell culture in serum-free media (Davami et al. 2014). Thanks to these combined attributes that solidified the position of CHO cells, they have become the gold-standard of the bioindustry and are responsible for the production of approximately 70% of all marketed RTPs (Pereira et al., 2018; Stolfa et al., 2018; Dziomba et al. 2023).

 

Fetal bovine serum (FBS) is commonly used as a supplement to basal media for in vitro cell cultures (including mammalian, animal, insect, and bacterial cells). Many studies have revealed its rich composition, containing essential macro- and micronutrients such as growth factors, adherent factors, amino acids, hormones, and vitamins (Michael Butler et al. 2020), which are crucial for proper cell function. In addition to the ethical issue, increasing demand, high cost, significant contamination risks (bacterial and viral), and batch-to-batch variability of FBS prompt us to seek alternatives. To reduce or eliminate FBS use, we have decided to implement culture media with various plant protein hydrolysates. Although serum-free media have been available for several decades, and their widespread adoption has increased, despite the fact that their compositions are undefined for users. Our goal is to make these serum-free media more accessible considering that serum-free media cost about 1.5 to 3 times more than serum-containing media.

 

Several publications highlighted the effects of vegetable proteins hydrolysate (peptones) as supplements in basal media, i.e : the enhancing of the cell growth and productivity of recombinant proteins (Burteau et al. 2003; Michael J. Betenbaugh et al. 2020). Some publications have shown an enhancing of the volumetric productivity more than 100%, and a 40% increase in cell number (Davami et al. 2014; Davami et al. 2015). Then, the composition of these plant hydrolysates varies according to the plant source and the production process. Thanks to their vegetable origins there are neither ethical issues, nor health standards.

 

In the present study, the effect of different peptone concentrations on the CHO-K1 cell line under various culture conditions (adherent/suspension) in a basal medium with a 10-fold reduction of FBS have been evaluated. Supplementation with wheat peptones at defined concentrations increased the viable cell density of CHO-K1 in adherent conditions by 190 – 265%.

 

Additionally, the suspension culture exhibited different results compared to the adherent conditions. In suspension cell culture, only three peptones – wheat, soy, and pea – enhanced viable cell density compared to the medium without hydrolysates. Actually, the only peptone supplementation is not enough to be completely serum-free, but that can reduce the FBS consumption.

Biography

Fabien Compeau is a researcher at both Organotechnie and Université Sorbonne Paris Nord, within the Tissue Engineering Research Unit. With a solid foundation in molecular and cellular biology. His current work centers on developing serum-free media. He completed three years of study at CY Cergy Paris Université and two years at Université Paris Cité. His internships provided valuable experience at the Institut de Recherche Criminelle de la Gendarmerie Nationale, the Institut Jacques Monod, and the Institut Pasteur, his research
focused on cell signaling, dynamics, and differentiation.

Igor Chourpa

Molecular optical spectroscopy as a PAT tool in USP bioprocessing

Abstract

Process analytical technology (PAT) aims at real-time, sampling-free monitoring of bioprocesses, for instance at the upstream stage (USP), when suspensions of CHO (chinese hamster ovary) cells grow in bioreactors and produce monoclonal antibodies (mAbs). The advantages of in-line PAT over the off-line approaches are numerous, from scientific, economic and ecological points of view. Using light as a non-invasive PAT tool is one of the promising ideas that give rise to various technological solutions. Nevertheless, the implementation of novel PAT approaches is limited and still requires further progress in analytical instrumentation and its integration in the whole process.
In this talk, we plan to present a short overview of advances in implementation of molecular optical spectroscopy as a PAT tool in upstream processing of therapeutic antibodies we obtained in the framework of our interdisciplinary research project CLIMB’IN. The project uses analytical spectroscopy based on light absorption and/or multi-angle scattering, both elastic (Rayleigh) and inelastic (Raman) for quantitative monitoring of multiple critical process parameters (CPPs). The advances reached by the CLIMB’IN consortium involve instrumental design and experimental protocols, with a particular focus made on machine learning algorithms. Brought together, these advances allow to improve the relevance and the robustness of the corresponding PAT approaches required for their industrial implementation.

Anne-Sophie GARY

Innovative tools to monitor biological variability of bioprocesses

Abstract

Today, virus-based products like oncolytic viruses and gene therapies are part of the most promising biotherapies. The development of biomanufacturing processes for viral products is not a straight road. It is generally associated with process complexity and biological variability. In depth expression system characterization by adding biological data to standard chemical data will help to understand bioprocesses. With a close collaboration between Naobios and Gensensor, case study from Vero cell substrate-based process will illustrate how transcriptomic data may help to understand and monitor biological variability linked to bioprocesses.

Biography

Anne-Sophie has a PhD in cell biology and is engineer in biotechnology. After working on cell stress and death pathways, Anne-Sophie specialized in the development of advanced therapy medicinal product. Now Principal Scientist at GenSensor, she is in charge of identifying biomarker panels on different expression systems and leads, with her team, the company’s internal research projects and scientific collaborations.