Shanghai, January 19, 2022 /PRNewswire/ -- Recently, Dr. Zhou Weichang, Chief Technology Officer and Executive Vice President of WuXi Biologics, published an opinion article in a professional journal, introducing the role of emerging technologies such as continuous production processes and disposable bioreactors in promoting the development of future biological processes, and sharing cutting-edge insights. The following are some exciting views:
Since the first monoclonal antibody (OKT3) was approved for marketing in 1986, the FDA has approved more than 100 antibody therapeutic products, including monoclonal antibodies (mAb), fusion proteins, antibody-conjugated drugs (ADCs) and bispecific antibodies (bsAbs).
Especially since 2014, the approved antibody drugs have shown explosive growth in the market for more than 70 products, including tumor necrosis factor-α (TNF-α), anti-PD-1 and anti-PD-L1 antibodies, and blockbuster drugs frequently appear. In 2021, the global sales of biological products are expected to exceed US$300 billion.12 In recent years, biological products have begun to dominate the global drug sales rankings. For example, in the first nine months of 2021, 13 of the 20 drugs with sales ranks among the top are biological products, including two mRNA new crown vaccines and 10 antibody products.
biological products have a highly complex structure and are usually produced by living cells. They require multiple process steps to purify their key features, namely key quality attributes (CQA), which can be different due to post-translational modifications that occur intracellularly or in subsequent production processes. Therefore, the process determines the product, and it can basically be said that "product is process" 3.
Looking back on the development of the biopharmaceutical industry over the past few decades, despite facing many technical and regulatory challenges, the industry's efforts in developing biologic liquid and preparation production processes and analytical methods have made great achievements, meeting a large number of clinical needs and saving the lives of many patients.
With the continuous emergence of innovative biopharmaceuticals, how to accelerate product launch while ensuring consistency in quality, especially in responding to the ever-changing COVID-19 pandemic, has put forward tests to the industry. The following challenges are promoting bioprocess innovation and bringing new directions to ensure product quality and stable supply in a cost-effective way:
Accelerate the process of product from concept, clinical trials to commercialization
Since the outbreak of the COVID-19 pandemic, the industry has responded quickly and accelerated the research and development and production of multiple vaccines (including two mRNA vaccines) and biopharmaceuticals (including multiple antibodies) through cooperation among multiple parties.
anti-COVID neutralizing antibody product development covers DNA sequence design, application for clinical trials of new drugs and approval for marketing. In order to speed up this process, the industry has adopted an integrated and transformative technical method without affecting product quality and safety: shortening the time for the product from DNA to IND application to 3 to 6 months, and completing the process of the COVID-19 neutralizing antibody from DNA to emergency use authorization (EUA) within 14 months. Thousands of kilograms of antibodies were produced in just a few months, which benefited patients around the world.4.
Recently, the development of new crown therapeutic drugs has reached an unprecedented speed, which may kick off the revolution in biological products research and development. Accelerating the development timeline has become one of the important features of future biotechnology development. In addition to continuing to apply innovative technologies to shorten R&D time, biotechnology development will also further focus on reducing production costs and a smaller "factory in the future" - comprehensively applying integrated continuous production processes, disposable bioreactors, and high digitization and automation, which will not only provide greater flexibility, higher output and more effective space utilization, but will also reduce production costs.
The changes brought by the COVID-19 epidemic to biological processes will also help the research and development of other more complex biological products, such as the knowledge and experience gained by the antibody-conjugated drugs and bispecific/multispecific antibodies industry from the rapid development of COVID-19 vaccines and biological products, and will also be more directly applied to the development of new vaccines and therapeutic biological drugs for other severe diseases. In the future, the research and development of new vaccines and biological drugs will break the tradition and will not take another 10 years.
Since the advent of therapeutic biological products, the selection strategies for cell lines for production have been continuously improved in order to improve production capacity and efficiency. These latest developments have allowed the industry to redevelop chemical, production and control (CMC) strategies and produce products that supply clinical trials in just three months. Codon optimization before DNA synthesis is a common method to achieve high protein expression levels. Its algorithm is based on dedicated codon and codon pairs using specific host cell lines, which may be more effective than algorithms based on using universal codon databases, helping to improve protein expression levels in specific host cell lines. In addition, since this strategy is effective for both transient transfection and stable transfection cell line expression systems, the optimization of customized codons also contributes to the entire R&D process.
As the CMC development process continues to accelerate over the years, toxicological research adopts protein materials obtained from cell populations has gained increasing recognition from regulatory agencies around the world. Therefore, it is crucial to select final clones with similar product CQA to proteins studied in toxicology. It is important to emphasize that instead of using peptide map analysis based on liquid synthesis and mass (LC-MS), cDNA based on next-generation sequencing (NGS) is used to quickly screen clones without any sequence variants in the cell population used in early development. During the cloning screening, cell line stability passage can be carried out simultaneously to further shorten the R&D time.
truncation or cleavage of recombinant proteins usually occurs during the development of complex biopharmaceutical molecules, which also poses challenges to downstream purification processes, and may also lead to a decrease in yield. Early stages of cell line selection are usually fed batches to screen out clones with less shear. This once again proves that a deep understanding of product and process progress in the early stages of R&D can help save a lot of time, cost and effort later.
More widely used continuous production processes and disposable bioreactors
In the future, continuous production processes will gradually and widely used various types of biopharmaceutical production such as monoclonal antibodies, dual antibodies, fusion proteins and recombinant proteins. This application also reflects the industry's development trend - meeting the growing demand for higher quality, higher yields and more accessible biological products.
In terms of upstream processes, perfusion culture has been widely used in clinical and commercial batch production. Compared with flow-added culture, perfusion culture has significant advantages in yield, quality, flexibility and cost-effectiveness. Advanced perfusion culture systems - such as the ultra-high-efficiency continuous production technology platform (WuXiUP™) independently developed by WuXi Biologics - can increase the cell density and yield in the production of almost all types of biologic drugs by 5 to 10 times compared with flow-added culture. In addition, continuous harvesting can shorten the residence time of the product in the bioreactor, thereby improving product quality.
WuXi Bio's ultra-high efficiency continuous production technology platform WuXiUP(TM)
With the advancement of technologies such as single-use bioreactors, flow-through chromatography and unidirectional tangential flow filtration, continuous production technology has successfully achieved continuous downstream processes in small or pilot scales. However, the investment in large-scale applications of the entire industry has lagged slightly.
uses automated steady-state perfusion culture, continuous harvesting without cells and extended product harvesting cycles, which has made great progress in product expression levels in continuous production upstream processes. Accordingly, in large-scale production, the output of continuous production downstream processes is also necessary to increase 5.
Technical Requirements for Human Use International Coordination Council (ICH) has issued a draft of Guiding Principles (Q13) on the continuous production of stock liquids and preparations (Q13) 6, which is currently being reviewed. The industry generally believes that the draft will encourage the application of continuous production processes in the research and development and production of biological products, and the continuous production process depends largely on partial or fully integrated downstream process unit operations.
For these two continuous production downstream process solutions, a foreseeable bottleneck is the lack of cost-effective ready-made or customized process analysis technology (PAT) tools and automated systems for process monitoring and real-time control. With the collaborative efforts of instrument and automation control solution suppliers, academia, engineers and the entire biopharmaceutical industry, this technology bottleneck is expected to be breakthroughs in the near future, thus allowing the full integrated continuous production downstream process to be widely used. The development strategy of
cell line is becoming increasingly advanced, and the cell culture medium is constantly optimized. Therefore, the expression level of cell culture products has been continuously improved, and the demand for bioreactors with a smaller area has also increased. In addition, continuous production processes will further promote the popularization of single-use bioreactors.
expands the use of single-use technology in other link operations, such as the introduction of a single-use tangential flow deep filtration system and a single-use ion exchange membrane chromatography device, which can achieve a complete single-use production process. The single-use technology also makes it possible to design new designs in production plants, such as the use of modular production units, thereby providing flexible production capacity, shortening product time to market, flexible conversion between different products, and also minimizing cross-contamination.
At present, enhanced continuous production technology has become a significant development trend, which will hopefully increase the output achieved by using these technologies. The complete continuous production process can be achieved by integrating and coordinating different unit operation processes to minimize gap time and increase production capacity. Using continuous production processes will also reduce the footprint of equipment used in the production of large-scale biological products and reduce overall capital investment.
Other bioprocess advancements include the implementation of automatic feeding, automatic sampling and minimizing pipeline assembly in large-scale production processes to achieve larger-scale process control, resulting in more robust performance and product quality. Process analysis techniques (PATs) such as Raman spectroscopy, and other online detection methods in large-scale production, enabling real-time detection and control of important process parameters and performance.
Other new analytical technologies such as polypeptide-based multi-attribute method (MAM) have higher sensitivity and product selectivity compared to capillary electrophoresis (CE) and high performance liquid chromatography (HPLC). In addition, emerging technologies such as surface plasmon resonance (SPR) and biolayer interference (BLI) can enhance the workflow in product release.
has two main advantages: rapid decision-making and advanced process control Various PAT platforms have been developed in the industry to monitor product aggregation and fragmentation degrees in continuous bioprocessing processes online, and automatically control live cell density 8, which helps to develop better processes and speed up CMC development progress.
A new generation of antibody drug process development trend
Due to the complexity of molecular structure, the process development of new biopharmaceuticals such as dual-antibody and antibody-coupled drugs is challenging. With the rapid growth of global antibody-coupled drug R&D product lines, the industry's demand for fully integrated antibody-coupled drug technology platform services is becoming increasingly strong, to support the process development and production of bio-coupled drugs in the process from DNA to IND. The research and development and production of bio-coupled drugs not only requires these fully integrated antibody-coupled drug technology platforms, but also require related services such as antibodies, linkers and coupling technology. In addition to shortening the time from DNA to IND, the integrated antibody-coupled drug technology platform also has the advantages of reducing risks during the development process. It also reflects that R&D companies pay more attention to the robustness of coupling process and drug-antibody ratio (DAR) control.
coupling technology involves various linker mechanisms and different payloads, coupling chemicals, coupling sites and drug-antibody diversity has also increased the variability in the production process, complicating the purification process and the analysis process of antibody-coupled drug structure and titers In the past few years, antibody-coupled drug R&D companies have made significant progress in improving the developability, productivity and functional performance of these complex drugs, but in pushing these drug models to a more streamlined R&D process, there are still certain challenges in responding to these difficulties. In order to deal with these difficulties, manufacturers need to adopt higher-level analytical methods.
In the production process of dual-antibody, the related by-products produced in the process such as chain mismatch, chain expression imbalance and incomplete assembly pose a considerable challenge to the downstream purification process. To help manufacturers deal with by-products of different types and content levels, the researchers proposed a toolbox-based dual-antibody purification method 9.
In addition to downstream methods tailored for dual antibody, perfusion cell culture can also significantly improve the quality of dual antibody (e.g., by increasing the percentage of monomers, determined by Caliper or capillary isoelectric focus analysis). Therefore, perfusion cell culture is a better choice method than traditional fed batch culture.10 The downstream and upstream strategies for dual antibody development discussed in this article can also be applied to multispecific antibody development.
Although there are still relevant challenges in the development of biological products, especially new and complex drug molecules such as antibody-conjugated drugs and dual-antibody, the industry has responded to these challenges through new analytical tools and high-throughput matrix-driven experimental designs compared with many years ago. Thanks to the more comprehensive, integrated and single-source biotechnology empowerment platform represented by WuXi Biologics, as well as the relevant knowledge and experience in the development of Chinese biopharmaceuticals in the fight against the new crown epidemic, enterprises will soon achieve the development of cost-effective therapeutic biological drugs and vaccines.
Reference
1. Business Research Company. Global Pharmaceuticals Opportunities and Strategies Market Report. Available at: .
2. Research and Markets. Global Biologics Market Opportunities and Strategies Report 2020: COVID-19 Impact and Recovery—Forecast to 2023, 2025 2030; 2021. Available at: .
3. BIO. How do Drugs and Biologics Differ? .
4. Zhang Z, Chen J, Wang J, et al. Reshaping cell line development and CMC strategy for fast responses to pandemic outbreak. Biotechnol. Prog. 2021; 37(5): e3186.
5. Zhou H, Fang M, Zheng X, Zhou W. Improving an intense and integrated continuous bioprocess platform for biologics manufacturing. Biotechnol. Bioeng. 2021; 118: 3618–3623
6. European Medicines Agency. ICH guideline Q13 on continuous manufacturing of drug substances and drug products. Draft version endorsed on 27 July 2021 .
7. Liu Z, Zhang Z, Qin Y, et al. The application of Raman spectroscopy for monitoring product quality attributes in perfusion cell culture. Biochem. Eng. J. 2021; 173: 108064.
8. Chen G, Hu J, Qin Y, Zhou W. Viable cell density on-line auto-control in perfusion cell culture aided by in-situ Raman spectroscopy. Biochem. Eng. J. 2021; 172: 108063.
9. Li Y, Wang Y, Shen K, Zhou W. A roadmap for IgG-like bispecific antibody Purification. In: Matte A, ed. Approaches to the Purification, Analysis, and Characterization of Antibody-Based Therapeutics. Elsevier; 2020: 167–179.
10. Qin Y, Ma R, Li Y, et al. Productivity and quality improvement for a bispecific antibody through the application of intensified perfusion cell culture. Unpublished data, 2021.
