Three productivity advantages of switching to transposon-based integration.

The development of stable cell lines remains a critical step in the production of recombinant and therapeutic proteins. As demand for faster development and higher yields increase, researchers are re-evaluating the tools they use for stable cell line generation. While random integration has traditionally been the standard method, transposon-mediated technologies are now gaining attention by delivering notable improvements in efficiency, stability, and consistency—three core pillars of productivity in cell line development.

Random vs transposon-mediated integration

Traditional cell line development has long relied on random integration vectors, where transgenes integrate into the host genome through non-homologous recombination. This process is inherently random, resulting in variable integration sites, inconsistent transgene copy numbers, and heterogeneous gene expression across the cell population. As a result, researchers need to conduct extensive screening campaigns to identify high-producing and stable clones, increasing development timelines and resource requirements.

In contrast, transposon-based systems use engineered DNA elements—often referred to as “jumping genes”—to introduce and stably integrate genes of interest into the host cell genome with greater efficiency and control. These transposons are flanked by inverted terminal repeats (ITRs) that serve as recognition sites for transposase enzymes. When activated, the transposase binds to these ITRs, excises the transposon carrying the therapeutic gene, and facilitates its integration into the host genome. Although transposon-mediated insertion remains semi-targeted, many transposon systems preferentially target transcriptionally active regions of the genome. This results in more consistent expression levels, improved clone stability, and a higher percentage of productive cell lines.

1. Efficiency: faster development timelines

Transposon-based integration has shown significant improvements in both the speed and efficiency of cell line development workflows. In a study by Balasubramanian et al., transposase-driven CHO cell pools achieved up to a ninefold higher productivity than those generated through random integration.1 This approach also substantially accelerated development timelines, enabling the production of clinical material in less than five months—a significant improvement over the standard 7–9 months for traditional methods.¹

These efficiency gains are driven by multiple technical advantages inherent to transposon-mediated integration, including:

• Reduced screening requirements
• Shorter selection times
• Elimination of intermediate pooling steps
• Creation of more uniform cell populations.²

By streamlining development and reducing operational overheads, transposon-based systems support faster, more cost-effective biotherapeutic development.

2. Consistency: comparable quality between pools and clones

Traditional development workflows require time-consuming clonal selection to ensure product consistency—a process that can add months to development schedules. However, transposon-based technologies enable consistent product quality even in non-clonal cell populations.

This was demonstrated by Agostinetto and colleagues during the COVID-19 pandemic, when rapid development of therapeutic antibodies became critically important.³ Their team used non-clonal, transposase-derived CHO cell pools to manufacture a COVID-19 monoclonal antibody at both 200L and 2,000L scales under cGMP conditions. Despite skipping clonal selection, the antibodies produced at both manufacturing scales exhibited comparable critical quality attributes to traditional workflows. Analysis of upstream and downstream processes, coupled with analytical testing, confirmed that the final product met all pre-defined specifications and was suitable for clinical use.

This emergency case highlights how transposon-mediated methods can enable high product quality while potentially streamlining traditional clonal workflows. The low variability among pools and clones enables more efficient screening, which saves time and resources while still maintaining quality. This approach also derisks development by generating sufficient material at the pool level—with quality consistent to clones—that can be used for toxicology studies, downstream process optimization, formulation work, and other critical activities.

3. Stability: sustained performance over time

Long-term cell line stability represents an essential regulatory requirement and manufacturing consideration for biotherapeutic production. Regulatory agencies require the demonstration of cell lines maintaining consistent productivity and product quality attributes throughout the manufacturing process.

In a study by Huhn et al., the stability of 19 transposase-derived CHO clones was evaluated over extended culture periods by comparing key performance indicators between early and aged cultures.⁴ The researchers specifically monitored productivity levels and Mannose-5 content—a critical glycosylation quality attribute that can impact therapeutic efficacy and immunogenicity.

Results showed that transposase-derived clones maintained consistent productivity and glycosylation profiles throughout the aging process. In contrast, clones generated through random integration had greater variability in both productivity and Mannose-5 levels as they aged.

These findings demonstrate that transposon-mediated integration not only accelerates development but also supports the generation of cell lines with stable performance and consistent product quality necessary to meet regulatory requirements.

Conclusion

Transposon-based integration technologies deliver measurable benefits in efficiency, consistency, and stability—directly impacting the speed, quality, and reliability of biotherapeutic manufacturing. By enabling higher productivity, shortened development timelines, and enhanced stability profiles they represent an attractive alternative to conventional random integration.

As companies continue to pursue faster and more reliable pathways to clinical development, particularly for complex biologics and emerging modalities, the ability to rapidly generate high-producing, stable cell lines without compromising on quality will be a significant competitive advantage.

Revvity’s CHOSOURCE™ TnT platform is a two-component expression system designed for recombinant protein production in GS knockout CHO cells. It combines a TnT transposon vector and transposase mRNA, enabling fast, reliable, and stable gene integration. Explore the complete CHOSOURCE TnT solution to see how it supports streamlined development and robust performance from early-stage research to clinical manufacturing. Our experienced global team offers expert guidance and support to help you adopt transposon-based approaches tailored to your biotherapeutic development needs. Download our infographic to discover how transposon technology can accelerate your biotherapeutic development timeline.