A HTE + PAT Strategy for Accelerated Bio-process Development
Adoption of High-Throughput Experimentation (HTE) for upstream cell culture is driving innovation, encouraging the realization of novel biotherapeutic candidates, and accelerating the development of promising products from molecule to market. High-throughput bioreactor systems with integrated analytics have revolutionized lab-scale capabilities for cell culture development through automation and advanced process control. Expert utilization of such platforms helps accelerate the development of biologic drug candidates from early process definition right through to process characterization, informing process scale-up, and beyond.
From a manufacturing and regulatory perspective, confidence in the reliability and scalability of a process is directly related to how well the CPPs are understood. This is particularly true in the case of Biologics where the tools to interrogate and understand processes are not as well developed as in the small molecule space. A lack of proper process understanding and control often leads to inability to adequately scale a process, product quality drift and costly delays arising from batch failures and concerns of the regulatory authorities which need to be addressed.
Thus, a key challenge for both CMC and Regulatory professionals within the Pharma/Biotech industry, is to minimize (or eliminate) these issues.
With a view to addressing this challenge, and in line with our mission of accelerating the development of life-changing medicines for patients, the team at APC decided to re-think each stage of how promising Biologics are developed and manufactured. It was clear that the old ways of doing things were not fit-for-purpose and a radically different approach would be necessary. With this in mind, we devised, created and implemented innovative new strategies and technologies which could be applied across process development, scale-up and tech transfer.
Regarding Process Development, our strategy was to couple High-Throughput Experimentation (HTE) with various analytical, modeling and statistical tools as depicted in the schematic below.
Our rationale for this approach was driven by our "engineering-first" philosophy of using Process Analytical Technologies (PAT) to build up a comprehensive picture of the critical process parameters at play in any given process. This PAT-driven approach has been a cornerstone in our success on over 1,000 large and small projects to date. However, the ability to integrate PAT with multiple parallel reactors opened up the possibility of generating large volumes of highly valuable, data-rich experimental outputs in a short period of time which could then be used to inform the development of very accurate process models. Further elements (e.g. statistical tools such as MVDA) integrated into this workflow would also provide complementary insights on parameter inter-dependencies.
APC took the decision to invest in an AMBR250 system with capacity to run 24 independent bioreactors simultaneously each with individual control over critical process parameters including agitation, temperature, pH, feeding/supplementation, and gassing (both sparger and headspace), etc. Complementing the extended capacity, several key analytics systems were integrated into the AMBR250, including a ViCell XR for automated cell counting, and a Nova Flex-II for quantification of relevant substrates, metabolites, osmolality, offline pH, dissolved gasses, and minerals. Raman spectroscopy was also incorporated directly into the AMBR system, allowing on-line quantification of Raman spectra across all bioreactors.
In addition, offline analytics, APC’s LC-MS system are utilized for spent media analysis, glycan analysis, peptide mapping, or product-specific CQA quantification. In-house flow cytometry methods are employed for investigating cellular features including Golgi & ER activity, programmed cell death, and cell cycle analysis.
The benefit and utility of implementing an experimental workflow utilizing automated reactors with integrated analytics, is multi-faceted, with tangible advantages attained at all stages of the process development journey, from early process design through to detailed characterization. Using representative scale-down process models, extended value may be delivered through troubleshooting and optimization of commercial-scale processes. Having capacity for 24 simultaneous bioreactors allows sophisticated Design of Experiment (DoE) strategiesto be rapidly executed and assessed, often in a single experimental investigation, meaning a greater understanding of the CPP's and limits of failure can be obtained. Enlisting the AMBR250 platform for cell culture experimentation shifts the focus of upstream development scientists away from time consuming bioreactor setup and operation, towards data-centered and hypothesis-driven experimental design, execution, and analysis. With automated data generation and collection through the AMBR250, coupled with iAchieve (a collaborative data management platform for maintaining active communication and robust digital documentation) a plethora of data is readily available on-demand at the fingertips of process development scientists. Ultimately, the high-throughput platform empowers science-focused, accelerated process development, enabling fast-tracked delivery of novel therapeutics to patients, reducing development timelines from molecule through to market.