A new approach to Bioprocess Optimisation
6 Nov 2012 by Evoluted New Media
Commercial production of biologics requires accurate and timely monitoring – Frederic Portait says Nuclear Magnetic Resonance can do this and help meet stringent regulations to boot… When producing biologics on a commercial scale, accurate and perceptive methods of monitoring/analysis of the cell culture are vital. Bioprocess optimisation aims to develop stable, high yielding and scalable production processes and to optimise cell viability/density to ensure a high quality and safe product.
It is essential to understand and control cell growth conditions early on. The Quality-by-Design (QbD) principles notably includes the understanding and characterisation of a cell culture process at its early stage of development. The purpose is to identify raw material and process parameters which are the most influential to the end-product quality. Although high pressure liquid chromatography (HPLC) or NOVA profiler bioanalysis methods are traditionally used, nuclear magnetic resonance spectroscopy (NMR) can rapidly provide reliable concentrationreliable concentration of all components at any given time.
The Rationale For NMR in Culture Media Analysis FDA (Food and Drug Administration) regulations are driving the biotech industry towards increasing the level of control, monitoring in bioprocess and using more chemically defined media. This increases the need for innovative and multiplex technologies to characterise media and monitor media component concentrations.
A large amount of bioprocesses are based on non-defined cell culture media for example hydrolysates, yeastolates and serum. These make bioprocess optimisation and consistent cell culture performance complicated to reach. Moreover, serum may be contaminated, encouraging a move towards defined serum-free culture media for bioprocessing. The detection of critical parameters specific to a cell line and a biomanufacturing process in a culture media will reduce batch-to-batch variability and fulfil modern product safety requirements.
HPLC methods are widely used with applications focusing on profiling certain media components. However, they remain limited when dealing with profiling of different nutrient classes or metabolites, especially of volatile components, in non-defined media. New developments are extending NMR methods, allowing the rapid, accurate and multiplex analysis of culture media at any stage of the process.
The concentration profiles obtained can then be used in several different ways, including: • Designing and optimising the medium; • Understanding cell-line nutrient requirements and optimising feeding strategies in the bioreactor; • Profiling raw material; • Troubleshooting in performance variability from batch-to-batch.
NMR, Quality-by-Design and the FDA The Office of Biotechnology Products (OBP) at the FDA organiszed a pilot on the implementation of QbD in 2009. QbD is a high-end manufacturing concept that requires a thorough understanding of the product and the process of manufacture. This includes:; • Understanding of variability in raw materials; • Knowledge of the relationship between the cell-line and the production process; • The product's critical quality attributes (CQAs); • Knowledge of the relationship between CQAs and the products clinical properties.
The complexity of biologic products makes QbD a difficult process. However, NMR methods have a capability to analyse a large range (>50) of media components simultaneously and provide comprehensive data at chosen times during a process, these approaches are uniquely suited to support part of the effort.
NMR Monitoring for Manufacture of Biologics and Biosimilars
NMR could change the face of bioprocess development and monitoring because it provides access to component identity, plus quantitative data, rapidly and easily from a single analysis. Small amounts of sample culture media can be taken across the required timeline along the process, and screened to identify and quantify metabolites, feed components or eventual contaminants in the media. No complex calibration is required and even unknown compounds can be rapidly identified from their NMR signals. It is thus possible to gain an in-depth understanding of the processes.
The nature of the technology makes it highly suitable for batch-to-batch monitoring and allows comparison of results across a whole project. NMR can handle a broad concentration range with linear concentration response and has excellent limits of detection from the micromolar level. When combined with chemometric methods, it is well-suited to tackle raw material lot-to-lot variability in cell culture process.
NMR methods can be applied to a wide range of cell culture and fermentation types, from stem cell culture to bacterial and yeast fermentation, including the production of biologics by Chinese Hamster Ovarian cells (CHO). NMR is able to rationalise the requirements of each cell-line, allowing efficient culture medium design. The untargeted technology can also provide a chemical composition snapshot of cultures and help determine influencing factors.
With increased regulations on product and process quality, biopharmaceutical companies must increase their level of control and monitoring in bioprocessing activities. Along with recent FDA guidance for the characterisation of biologics and biosimilars, NMR shows more and more benefits in this field.
The oOverall aAdvantages of NMR mMonitoring • Rapidly provides access to accurate absolute concentration data • Identifies media components, including previously undefined nutrients and metabolites • Rapidly screens culture medium and provides profiling in combination with existing chemometric tools • Analyses multiple components • Has a good LOD from 1-10µM with a linearity over a broad concentration range • Provides detailed consumption profiles whatever the type of batch (straight, fed batches or perfusion batches) to identify variations • Deliver robust and reproducible quantitative analysis of complex media • Troubleshoot and solve bioprocess performance variability to reach consistent and optimal performance
Conclusion The contribution of NMR to the development and manufacturing of biologics is growing with the application of more sophisticated automation and analytical support technologies. NMR has proven itself highly capable of analysing complex culture media, and can now be used over a range of production platforms and cell systems to provide insight into the cell culture process and its development. Thus, NMR can support the development of a bioprocess in line with QbD. Results provide the information necessary to optimisze cell culture media making selection and design processes rapid and cost effective.
Author:
Dr FC Girard, CEO Spinnovation Analytical
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