Revolutionizing cancer treatment: The critical role of stable cell lines

Developments in stable cell line development have been central to several pivotal discoveries in cancer research, most notably advances in chimeric antigen receptor (CAR)-T cell therapy. Due to their uniformity and reproducibility, stable cell lines are indispensable instruments in pharmacological and biotechnological research.

This article examines the present and future potential of stable cell lines, exploring their significance and applications, with a specific focus on their use in oncological studies.

The multifaceted utility of stable cell lines

An array of recent scientific projects and applications have thoroughly demonstrated the wide-ranging application of stable cell lines. This technology has been especially prevalent in areas such as gene function studies, drug screening, and the production of recombinant proteins and antibodies.

Progressions in cell line development have been key to furthering advances in recombinant protein therapeutics. Improvements in the production timeline and the streamlining of high-yield cell line generation have the potential to considerably expedite the creation of both original therapeutic products and biosimilars.1

The stable integration of the transgene in the genome of an expression host offers a myriad of benefits for biopharmaceutical production. Two key attributes of stable cell lines are their ability to mitigate batch-to-batch discrepancies in recombinant protein production and their capacity to yield a greater degree of protein expression in contrast with methods involving transient transfection.

Mammalian cell lines have long been the primary system in the manufacture of biopharmaceuticals. This dominance has largely been due to their ability to synthesize proteins with post-translational modifications (PTMs) possessing similar characteristics to those naturally occurring in humans.2

Chinese hamster ovary (CHO)-derived cell lines remain the most widely used platform for therapeutic protein production, such as CHO-DXB11 and CHO-DG443.

This has remained the case despite the availability of several mammalian systems on the market, with approximately 70% of all recombinant biologics and almost 80% of approved monoclonal antibodies being produced in CHO systems.3,4

Sino Biological boasts a rich legacy of cell line development spanning over 15 years. The company offers custom-tailored CHO stable cell line development services for protein and antibody production (Figure 1).

Workflow of Stable Cell Line Development for Protein and Antibody Production at Sino Biological.

Figure 1. Workflow of Stable Cell Line Development for Protein and Antibody Production at Sino Biological. Image Credit: Sino Biological Inc.

Stable cell lines continue to be the most popular option for biopharmaceutical production and provide important research tools for applications such as compound screening, assay development, and target discovery and validation.

Stable cell lines generated to overexpress specific proteins continuously are a commonly used tool in assay applications, but generating these overexpression cell lines is a time-consuming and complicated process.

Sino Biological leverages its extensive experience to offer one-stop stable cell line development services, enabling a wide range of basic research and drug discovery (Figure 2), including the development of overexpression stable cell lines with difficult-to-transfect cells and high expression of difficult-to-express proteins.

Flowchart of Overexpression Stable Cell Line Generation for Assay Applications at Sino Biological.

Figure 2. Flowchart of Overexpression Stable Cell Line Generation for Assay Applications at Sino Biological. Image Credit: Sino Biological Inc.

CAR-T cell therapy and the need for stable cell lines

CAR-T cell therapy is at the forefront of a new era in oncological treatment. Stable cell lines are a key contributor to the success of this method, not just in the foundational development of CAR-T cell therapies but also in their ongoing optimization and quality control.

CAR T-cell therapy was brought into clinical practice in 2017 and has since become integral to treating lymphoid malignancies (primarily those of B-cell origin), yielding impressive therapeutic results.5 These conditions include non-Hodgkin lymphoma, lymphoblastic leukemia, and plasma cell myeloma.

This pioneering treatment modality, which markedly departs from traditional cancer therapies, reengineers a patient's T cells to recognize and combat cancer cells (Figure 3).

CAR-T Cell Therapy.

Figure 3. CAR-T Cell Therapy. Image Credit: Sino Biological Inc.

As a highly personalized immunotherapy, CAR-T cell therapy has transformed cancer treatment, but its development is challenging.

Consistent reproducibility is necessary to optimize the specificity and efficacy of CAR-T cell therapy. Stable cell lines can address this particular developmental challenge.

Stable cell lines pave the way for more effective, robust, and reliable CAR-T therapies. CAR-positive samples are critical to developing CAR-T cell assays, while stably transfected cell lines (Jurkat and CHO) have been employed as a viable source of positive controls for CAR-T cell assays.6

CAR-transduced cell lines represent an ideal option for establishing homogenous controls designed to express known levels of the CAR construct in early antibody screening and assay development.

These stable cell lines contain limited T-cell markers required to create gating approaches for patient specimen analysis. They are a robust choice for early reagent screening due to the difficulties of sourcing an appropriate number of ‘control’ CAR-T cells used for CAR-T measurement.

Overexpression-stable cell lines genetically modified to overexpress specific tumor antigens are suitable in vitro cell models for CAR-T cell therapy. They are well suited to evaluating CAR-T cells’ specificity and cytotoxicity.

For example, CAR-T cells' specific killing activity and cytokine secretion against target cells can be measured through in vitro co-culture of these target-expressing cell lines with CAR-T cells.7

These cell lines are central to CAR-T therapy's continuous refinement and enhancement. Assay data acquired via these in vitro assessments is key to efficiently evaluating and optimizing the CAR-T therapy development process.

Conclusion

The cancer research and treatment field continues to expand, with stable cell lines representing a cornerstone technique that underpins many historic advancements that have revolutionized oncology.

Progress in CAR-T therapy, biopharmaceutical manufacturing, and other research fields showcase their indispensable contribution.

With any dynamic scientific field, there remains a need for ongoing innovation and optimization. This is also the case in stable cell line development, with the optimization of cell culture conditions, advancing cell line engineering technologies, and introducing emerging screening technologies.

Stable cell lines must be specifically developed for the project in question. By providing a holistic service suite, Sino Biological can efficiently construct custom stable cell lines with enhanced stability, improved productivity, and a quick turnaround.

This streamlined process facilitates the functional verification of CAR-T cells and caters to a wide range of other research and production requirements.

References and further reading

  1. Lai T, Yang Y, Ng SK. Advances in Mammalian cell line development technologies for recombinant protein production. Pharmaceuticals (Basel). 2013;6(5):579-603. Published 2013 Apr 26. doi:10.3390/ph6050579
  2. Zhu J. Mammalian cell protein expression for biopharmaceutical production. Biotechnol Adv. 2012;30(5):1158-1170. doi:10.1016/j.biotechadv.2011.08.022
  3. Zhu MM, Mollet M, Hubert RS, Kyung YS, Zhang GG. Industrial Production of Therapeutic Proteins: Cell Lines, Cell Culture, and Purification. Handbook of Industrial Chemistry and Biotechnology. 2017;1639-1669. Published 2017 May 3. doi:10.1007/978-3-319-52287-6_29
  4. Xu WJ, Lin Y, Mi CL, Pang JY, Wang TY. Progress in fed-batch culture for recombinant protein production in CHO cells. Appl Microbiol Biotechnol. 2023;107(4):1063-1075. doi:10.1007/s00253-022-12342-x
  5. Ramesh P, Hui HYL, Brownrigg LM, Fuller KA, Erber WN. Chimeric antigen receptor T-cells: Properties, production, and quality control. Int J Lab Hematol. 2023;45(4):425-435. doi:10.1111/ijlh.14121
  6. Sarikonda G, Mathieu M, Natalia M, et al. Best practices for the development, analytical validation and clinical implementation of flow cytometric methods for chimeric antigen receptor T cell analyses. Cytometry B Clin Cytom. 2021;100(1):79-91. doi:10.1002/cyto.b.21985
  7. Golubovskaya V, Zhou H, Li F, et al. Novel CD37, Humanized CD37 and Bi-Specific Humanized CD37-CD19 CAR-T Cells Specifically Target Lymphoma. Cancers (Basel). 2021;13(5):981. Published 2021 Feb 26. doi:10.3390/cancers13050981

Acknowledgments

Produced from materials originally authored by Sino Biological. 

About Sino Biological Inc.

Sino Biological is an international reagent supplier and service provider. The company specializes in recombinant protein production and antibody development. All of Sino Biological's products are independently developed and produced, including recombinant proteins, antibodies and cDNA clones. Sino Biological is the researchers' one-stop technical services shop for the advanced technology platforms they need to make advancements. In addition, Sino Biological offer pharmaceutical companies and biotechnology firms pre-clinical production technology services for hundreds of monoclonal antibody drug candidates.

Sino Biological's core business

Sino Biological is committed to providing high-quality recombinant protein and antibody reagents and to being a one-stop technical services shop for life science researchers around the world. All of our products are independently developed and produced. In addition, we offer pharmaceutical companies and biotechnology firms pre-clinical production technology services for hundreds of monoclonal antibody drug candidates. Our product quality control indicators meet rigorous requirements for clinical use samples. It takes only a few weeks for us to produce 1 to 30 grams of purified monoclonal antibody from gene sequencing.


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Last updated: Jul 8, 2024 at 6:18 AM

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