Viral production pipelines have suffered from a lack of scalability. Researchers are increasingly shifting from adherent cell culture to suspension culture, which allows for greater scalability and brings speed and cost efficiencies to viral vector production. This move also reduces the chances of batch-to-batch variation and brings viral vector production processes more in line with other large-scale production processes like those used for monoclonal antibody production (Ansorge et al., 2019).
Automation carries several advantages wherever it is implemented across viral vector production workflows. By removing a significant amount of risk due to human error and variation, automation helps to streamline production and promotes reproducibility. Automation in liquid handling enables more reliable single-cell seeding, which reduces the chance of contamination from other clones and microbes. Automated cell culture with real-time monitoring ensures cells are maintained at optimal growing conditions, facilitating faster scaling up and less risk to workflows (Holland & Davies, 2020).
The
C.STATION is CYTENA’s all-in-one platform for automated cell line development workflows. It houses several best-in-class instruments, including the
UP.SIGHT to accelerate the generation of cell lines for viral vector production while enhancing quality and ensuring regulatory alignment (Fig. 2).
Figure 2. The C.STATION combines automated cell dispensing, liquid handling, cell culture, and more to provide the ultimate solution for cell line development workflows.
Challenges and Future Perspectives
The increased demand for AAV-based viral vectors for gene therapies has highlighted the need for faster, more cost-efficient production methods paired with heightened safety and regulatory compliance. Perennial issues, such as immunogenicity and targeting of viral vectors to appropriate tissues, limit their translational impact. Advances in targeting promise to increase the specificity of viral vectors for specific tissues and cell types (Srivastava et al., 2021; Zhang et al., 2016), while methods including capsid modification with polyethylene glycol (PEG) are helping to reduce the immunogenicity of gene therapy vectors (Kreppel & Hagedorn, 2021).
Regulatory bodies require proof of clonality for cell lines used to produce therapeutics, which is challenging to prove using manual cell seeding methods. The UP.SIGHT single-cell dispenser and dual-imager from CYTENA images cells as they are dispensed and after they settle in the well, making it simple to prove clonality to regulators (Fig. 3).
Figure 3. The UP.SIGHT enables >97% single-cell dispensing efficiency, significantly derisking production pipelines while eliminating the need for manual input.
Conclusion
Viral vector production stands at the forefront of therapeutic innovation, enabling groundbreaking advances in gene therapy and vaccine development. CYTENA’s UP.SIGHT technology supports these efforts by enabling regulatory compliance through accurate single-cell seeding. As demand for viral vectors grows, ongoing technological advancements will be essential in meeting safety, scalability, and precision requirements, bringing promising treatments closer to patients worldwide.
CYTENA drives the future of viral vector production by enabling researchers to streamline their processes while ensuring regulatory compliance.
References
- Ansorge, S., Burnham, M., Kelly, M., McDermott, R., & Jones, P. (2019). Scale-up considerations for improved yield in upstream viral vector production. Cell and Gene Therapy Insights, 5(12), 1719–1725.
- Bulcha, J. T., Wang, Y., Ma, H., Tai, P. W. L., & Gao, G. (2021). Viral vector platforms within the gene therapy landscape. Signal Transduction and Targeted Therapy, 6(1), 53.
- Finer, M., & Glorioso, J. (2017). A brief account of viral vectors and their promise for gene therapy. Gene Therapy, 24(1), 1–2.
- Gimpel, A. L., Katsikis, G., Sha, S., Maloney, A. J., Hong, M. S., Nguyen, T. N. T., Wolfrum, J., Springs, S. L., Sinskey, A. J., Manalis, S. R., Barone, P. W., & Braatz, R. D. (2021). Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies. Molecular Therapy. Methods & Clinical Development, 20, 740–754.
- Hasanzadeh, A., Hamblin, M. R., Kiani, J., Noori, H., Hardie, J. M., Karimi, M., & Shafiee, H. (2022). Could artificial intelligence revolutionize the development of nanovectors for gene therapy and mRNA vaccines? Nano Today, 47, 101665.
- Holland, I., & Davies, J. A. (2020). Automation in the Life Science Research Laboratory. Frontiers in Bioengineering and Biotechnology, 8, 571777.
- Kreppel, F., & Hagedorn, C. (2021). Capsid and Genome Modification Strategies to Reduce the Immunogenicity of Adenoviral Vectors. International Journal of Molecular Sciences, 22(5), 2417.
- Lei, T., Wang, Y., Zhang, Y., Yang, Y., Cao, J., Huang, J., Chen, J., Chen, H., Zhang, J., Wang, L., Xu, X., Gale, R. P., & Wang, L. (2024). Leveraging CRISPR gene editing technology to optimize the efficacy, safety and accessibility of CAR T-cell therapy. Leukemia.
- Srivastava, A., Mallela, K. M. G., Deorkar, N., & Brophy, G. (2021). Manufacturing Challenges and Rational Formulation Development for AAV Viral Vectors. Journal of Pharmaceutical Sciences, 110(7), 2609–2624.
- Travieso, T., Li, J., Mahesh, S., Mello, J. D. F. R. E., & Blasi, M. (2022). The use of viral vectors in vaccine development. Npj Vaccines, 7(1), 75.
- Wang, J.-H., Gessler, D. J., Zhan, W., Gallagher, T. L., & Gao, G. (2024). Adeno-associated virus as a delivery vector for gene therapy of human diseases. Signal Transduction and Targeted Therapy, 9(1), 78.
- Zhang, C., Yao, T., Zheng, Y., Li, Z., Zhang, Q., Zhang, L., & Zhou, D. (2016). Development of next generation adeno-associated viral vectors capable of selective tropism and efficient gene delivery. Biomaterials, 80, 134–145.