How to generate stable cell lines?

Overview of Stable Cell Lines

Historical Context

The development of stable cell lines traces back to the mid-20th century when advances in cell culture and molecular biology paved the way for genetic engineering. One of the pioneering efforts was the establishment of the HeLa cell line in 1951, the first human cell line to be successfully cultured indefinitely. While HeLa cells were not originally engineered to express a transgene, their use in research demonstrated the potential of immortalized cell lines for continuous scientific study.

In the 1970s, with the advent of recombinant DNA technology, scientists began to explore methods for stably integrating foreign genes into the genomes of mammalian cells. One of the early breakthroughs was the development of the Chinese Hamster Ovary (CHO) cell line as a host for the production of recombinant proteins. The CHO cell line, established in 1957, became a workhorse in biotechnology, especially after the first successful production of recombinant tissue plasminogen activator (tPA) using CHO cells in the 1980s. This success demonstrated the feasibility of using genetically engineered stable cell lines for large-scale protein production, marking the beginning of their widespread use in both research and industry.

Today, the creation of stable cell lines involves a series of intricate steps, including the selection of an appropriate host cell line, vector design, transfection, selection, and cloning. Each of these steps requires careful consideration and optimization to achieve a cell line that not only expresses the transgene stably but also meets the specific needs of the intended application, such as high-yield protein production or precise gene function studies.

What Are Stable Cell Lines?

Definition and Role

Stable cell lines are clonal populations of cells that have been genetically engineered to permanently integrate a foreign gene, known as a transgene, into their genome. This integration is achieved through gene editing technologies, which ensure that the transgene is stably inherited and expressed in all daughter cells during cell division. The transgene typically encodes a molecule of interest, such as a therapeutic protein (monoclonal antibodies, insulin…), a reporter protein (GFP, luciferase…), or a gene regulator (siRNA, CRISPR/Cas9 components…). Unlike transient expression systems, where gene expression is temporary, stable cell lines provide long-term and continuous production of the desired protein, making them invaluable for applications that require sustained expression.

Characteristics of stable cell lines

Stable cell lines exhibit several key characteristics that distinguish them from other expression systems:

• Genetic Stability: The transgene is permanently integrated into the cell’s genome, ensuring its transmission to progeny cells during mitosis. This genetic stability is crucial for applications that require consistent gene expression over extended periods.
• Consistent Expression: Stable cell lines are engineered to provide uniform transgene expression, which is essential for reproducibility in experimental settings and consistency in manufacturing processes. This consistency is achieved through the careful selection and cloning of cells that meet the desired expression criteria.
• Longevity: Unlike transiently transfected cells, which express the transgene only briefly, stable cell lines are capable of maintaining gene expression over prolonged periods, often spanning several months or years. This longevity is particularly advantageous for long-term studies and industrial-scale production.
• Scalability: Stable cell lines can be readily expanded and scaled up, enabling the production of large quantities of the target protein or facilitating high-throughput screening. This scalability makes them highly suitable for both research and commercial applications.
• Reproducibility: Derived from a single cell clone, stable cell lines offer a high degree of reproducibility, ensuring consistent results across experiments and production batches. This reproducibility is critical for the validation and standardization of scientific and industrial processes.

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Comparison with transient cell lines

The primary distinction between stable and transient cell lines primarily lies in the duration and reliability of gene expression:

• Transient Cell Lines: In transient transfection, the transgene is introduced into cells without genomic integration. As a result, gene expression is short-lived, typically lasting only a few days before the plasmid DNA is degraded or diluted during cell division. Transient cell lines are suitable for short-term studies or rapid expression needs. However, their temporary nature limits their utility in long-term experiments or large-scale production.

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• Stable Cell Lines: In contrast, stable cell lines integrate the transgene into the host genome or maintain it as an episomal plasmid, allowing for continuous and long-term gene expression. This integration makes stable cell lines the preferred choice for applications requiring sustained expression, such as biomanufacturing or chronic studies. While the development of stable cell lines is more time-consuming and technically demanding than transient transfection, the benefits of sustained and reliable gene expression often justify the investment.

Why Generate Stable Cell Lines?

Importance and Benefits of using stable cell lines in research and industry

Stable cell lines play a crucial role in advancing scientific research and driving industrial innovation. Their ability to provide consistent and prolonged gene expression makes them indispensable for a range of applications, from basic research to large-scale bioproduction.

Enhanced Research Precision and Consistency

Stable cell lines are essential for experiments that demand long-term study and reproducibility. In molecular biology and genetics, the ability to study gene function over extended periods is vital for understanding complex biological processes. Stable cell lines offer the precision needed to explore these processes, allowing researchers to control for variables and ensure that the observed effects are due to the gene of interest rather than fluctuations in gene expression. This level of control and consistency is key to generating reliable data, which is fundamental for advancing scientific knowledge.

Industrial Scalability and Efficiency

In the biotechnology and pharmaceutical industries, stable cell lines are the backbone of commercial production processes. They enable the large-scale production of biologics, such as recombinant antibodies, hormones, and vaccines, by providing a robust and scalable system for continuous protein expression. The stability of these cell lines ensures that each production batch meets stringent quality standards, reducing variability and increasing efficiency. This scalability is crucial for meeting global demand for therapeutic proteins and vaccines, especially during public health emergencies.

Cost-Effectiveness in Long-Term Studies

While the development of stable cell lines requires an initial investment of time and resources, their long-term benefits far outweigh the costs. Once established, stable cell lines can be maintained and used for several months, providing a reliable source of gene expression without the need for repeated transfection or complex re-engineering. This cost-effectiveness is particularly advantageous in drug development, where sustained expression is required for extended preclinical studies, high-throughput screening, and the production of clinical-grade materials.

Versatility Across Multiple Applications

Stable cell lines are versatile tools that can be tailored to meet the specific needs of various research and industrial applications. They are not only used for the production of therapeutic proteins but also play a critical role in gene editing, functional genomics, and toxicology studies. For example, in gene editing, stable cell lines can be engineered to express components of CRISPR/Cas9 systems, enabling precise genetic modifications that are propagated across cell generations. This versatility makes stable cell lines an invaluable resource across multiple domains, from basic research to applied biotechnology.

Regulatory Compliance and Product Safety

In the context of biopharmaceutical production, stable cell lines offer a level of consistency and control that is essential for meeting regulatory requirements. Regulatory agencies, such as the FDA and EMA, require that biologics produced for therapeutic use are manufactured in a consistent and reproducible manner. Stable cell lines ensure that the production process is reliable, with minimal batch-to-batch variability, which is critical for ensuring the safety and efficacy of the final product. This compliance with regulatory standards is a key benefit of using stable cell lines in the biomanufacturing industry.

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Key applications

• Antibody Production: In the production of therapeutic antibodies, stable cell lines, particularly those derived from CHO (Chinese Hamster Ovary) cells, are the industry standard. These cell lines are engineered to produce high yields of monoclonal antibodies, which are used to treat a wide range of diseases, including cancers, autoimmune disorders, and infectious diseases. The stability and scalability of these cell lines make them ideal for the large-scale production required in commercial biomanufacturing. Beyond industrial applications, stable cell lines are widely used in academic settings for the production of recombinant proteins, which are then used in various biochemical assays, structural biology studies, and other research applications.
• Gene Editing and Functional Studies: Stable cell lines are essential for gene editing applications, such as those involving CRISPR/Cas9 technology. By integrating components like guide RNAs and Cas9 proteins into the genome, stable cell lines can be created to perform precise gene knockouts or knock-ins. These cell lines are valuable tools for studying gene function, investigating disease mechanisms, and developing gene therapies.
• Drug Development: Stable cell lines are critical in the drug discovery and development process. They are used to create cell-based assays that enable the screening of thousands of compounds for potential therapeutic effects. For instance, cell lines expressing specific receptors or enzymes can be used to identify small molecules that modulate these targets, providing early-stage data on drug efficacy and toxicity.
• Vaccine Development: Stable cell lines are also employed in the production of viral proteins used as antigens in vaccines. For example, cell lines can be engineered to produce large quantities of viral surface proteins, which are then purified and used to formulate vaccines. This application is particularly important in the rapid development of vaccines for emerging infectious diseases.

Step-by-Step Guide to Generating Stable Cell Lines

Generating a stable cell line is a complex and multi-step process that requires careful planning and execution. The success of generating stable cell lines depends on several critical factors, each of which plays a pivotal role in ensuring the stable and consistent expression of the desired transgene.

General Workflow

Vector Design and Construction

• Selection of the appropriate vector type: This decision is critical and must be based on the specific experimental requirements and cell type being used. For example, plasmid vectors are often preferred for their ease of use and versatility, while viral vectors may be preferred for their high transfection efficiency, especially in difficult-to-transfect cells.
• Design the expression cassette: This involves incorporating several key components, discussed below, that together ensure robust gene expression. The promoter, which initiates transcription, must be carefully selected to match the desired expression profile, whether constitutive, inducible, or tissue-specific. In addition to the promoter, the inclusion of a selectable marker gene is essential to facilitate the identification of successfully transfected cells. In addition, sequences such as the polyadenylation signal and enhancers are incorporated to stabilize the mRNA and increase gene expression, respectively.
• Gene cloning and optimization : This step involves precise insertion of the gene into the vector, with attention to optimizing codon usage to match the host cell’s preferences. Codon optimization can significantly enhance the translation efficiency and, consequently, the protein expression levels. Once the gene is successfully cloned, it is critical to verify the integrity of the construct through sequencing, ensuring that the gene is correctly inserted and that all components are functional.

To learn more about strategies for optimizing vector design and gene integration, read our blog “Gene Integration in Antibody Production: Overcoming Challenges for Optimal Clone Selection”.

• Pilot testing : This step involves a transient transfection to assess the efficiency of gene expression. The results of this pilot test provide valuable insights into the performance of the vector and highlight any necessary adjustments that need to be made before proceeding to stable transfection. This iterative process of testing and optimization is key to ensuring that the final vector design will lead to the successful generation of a stable cell line with the desired characteristics.

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Transfection

• Selection of the transfection method: This step depends on the specific cell type being used and the experimental requirements. Common transfection methods include lipofection, electroporation, and calcium phosphate precipitation, each with its advantages and limitations. For example, lipofection is often chosen because of its ease of use and high efficiency in many mammalian cell lines, while electroporation is preferred for difficult-to-transfect cells because of its ability to deliver DNA directly into the nucleus using electrical pulses. The choice of method has a significant impact on the overall success of transfection, so it is important to match the method to the characteristics of the target cells.
• Optimizing the transfection conditions: This involves fine-tuning several parameters, including DNA concentration, reagent ratios, and incubation times. The goal is to maximize transfection efficiency – ensuring that a high percentage of cells are correctly transfected – while minimizing cytotoxicity, which can lead to cell death and reduce the number of viable transfected cells. Each cell type may require specific adjustments to these parameters, and achieving the right balance is critical to the success of the transfection process. Iterative testing and refinement are often required to determine the optimal conditions that yield the highest transfection efficiency with minimal adverse effects on cell viability.
• Stable transfection: Once optimal transfection conditions are established, the focus shifts to achieving stable transfection, where the vector is introduced under conditions conducive to its stable integration into the host cell genome.

Selection and Screening of Stable Clones

These selection steps ensure that only cells with stable and robust integration of the transgene are retained for further use.

• Selective pressure: This step is designed to eliminate non-transfected cells by incorporating a selectable marker gene into the vector that confers resistance to an antibiotic. This selective pressure is essential to enrich the cell population with the desired genetic modification, thereby increasing the likelihood of isolating stable clones.
• Single-cell cloning: Individual cells are isolated to generate monoclonal populations. This can be accomplished by techniques such as Limiting Dilution, Fluorescence-Activated Cell Sorting (FACS), or more sophisticated technologies like Verified In-Situ Plate Seeding (VIPS). Single-cell cloning is a pivotal step because it ensures that the resulting clones are derived from a single progenitor cell, leading to a genetically homogeneous population. This homogeneity is crucial for reproducibility in subsequent experiments and applications.

At ProteoGenix we harness the power of cutting-edge for VIPS technology</ to ensure precision and reliability in your cell line development:

• Precision in single-cell seeding
• Real-time image verification
• Accelerated development timelines
• Robust regulatory compliance

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• High-producing clones screening: The goal is to identify clones with the highest gene expression levels. Various assays are employed to quantify the expression of the gene of interest, with techniques such as enzyme-linked immunosorbent assay (ELISA), quantitative PCR (qPCR), and flow cytometry being commonly used. ELISA is particularly useful for detecting and quantifying secreted proteins, while qPCR provides precise quantification of mRNA levels, and flow cytometry allows for the analysis of protein expression at the single-cell level.
• Genetic characterization: This step involves confirming the integration of the transgene and assessing the stability of its expression over time. Techniques such as Southern blotting and qPCR are commonly employed for this purpose. Southern blotting provides information on the copy number and integration sites of the transgene, ensuring that the genetic modification is as intended. qPCR, on the other hand, can be used to monitor the consistency of gene expression across multiple passages, verifying that the expression remains stable over extended culture periods.

Expansion and Validation of Stable Clones

• Expansion of clones: Once high-producing clones have been identified, they are gradually expanded from small-scale cultures to larger volumes. This expansion must be carried out carefully, with continuous monitoring of cell growth and expression levels to ensure that the clones maintain their productivity during scale-up. Monitoring is crucial because changes in culture conditions, such as nutrient availability or cell density, can affect gene expression levels and cell viability.
• Stability testing : Stability testing involves assessing the long-term consistency of gene expression and cell viability over multiple cell passages. This step is vital because even clones that initially show high expression levels can sometimes lose their productivity over time, particularly if the transgene is not stably integrated or if the cells undergo genetic drift. Stability testing typically involves serially passaging the clones and periodically measuring gene expression levels, often using techniques like quantitative PCR (qPCR) or flow cytometry. By confirming that gene expression remains stable over extended culture periods, researchers can ensure that the clones will continue to perform reliably in large-scale applications.
• Functional validation: While high expression levels are important, it is equally crucial to confirm that the expressed antibody is functionally active. Functional validation involves conducting relevant biological assays to assess the activity of the expressed protein. For example, if the gene of interest encodes an enzyme, enzymatic activity assays would be used to measure the catalytic function of the protein. Alternatively, for proteins involved in binding interactions, such as antibodies or receptors, binding assays could be employed to evaluate the affinity and specificity of the protein for its target.

Scale-Up and Production

• Optimization of culture conditions: As the scale of production increases, so too does the complexity of the culture environment. The goal of this optimization process is to replicate the conditions that were successful at smaller scales while adapting them to the challenges of large-scale production. Key factors that need to be optimized include media composition, oxygen levels, and bioreactor settings. Learn more about how to optimize culture conditions for large-scale production, read our blog: “From Lab Bench to Bioreactor: Overcoming Scale-Up Challenges in Antibody Production”.
• Pilot production: These small-scale runs serve as a test bed for the scaled-up production process, providing an opportunity to troubleshoot and optimize before committing to full-scale production.
• Large-scale production: At this stage, the stable cell line is cultured in large bioreactors designed to produce the target protein at the required scale. This involves maintaining strict control over all aspects of the production process, including culture conditions, bioreactor operation, and downstream processing.

Quality Control and Regulatory Compliance

• Quality control (QC): Quality control testing is not just a final step but an ongoing process that continues throughout the production cycle to monitor consistency and maintain high standards. This involves conducting a series of rigorous tests designed to assess the purity, potency, and safety of the final product.
• Purity testing ensures that the product is free from contaminants, such as residual host cell proteins, DNA, or adventitious agents, which could compromise its quality or safety.
• Potency testing is performed to confirm that the product maintains its expected biological activity, which is essential for its effectiveness in therapeutic or research applications.
• Safety testing, on the other hand, focuses on detecting any potential risks associated with the product, including toxicity or immunogenicity.
• Regulatory documentation: In tandem with QC must be meticulously prepared to comply with the relevant regulatory requirements. This documentation is essential for obtaining approval from regulatory bodies, such as the U.S. Food and Drug Administration (FDA) or the European Medicines Agency (EMA), for clinical or commercial use. Key components of this documentation include cell line authentication, which verifies the identity and genetic integrity of the cell line, and characterization data, which provides detailed information about the genetic modification, expression levels, and stability of the cell line.

Best Practices for Successful Stable Cell Line Generation

Vector Types

Vectors are essential tools in molecular biology that facilitate the delivery and expression of genes in host cells. When generating stable cell lines, the choice of vector is critical, as it determines the efficiency of gene integration and expression. There are several types of vectors that researchers can use:

• Plasmid Vectors: Circular DNA molecules that replicate independently within the host cell. They are easy to manipulate and are commonly used for gene cloning and expression in bacteria, but can also be adapted for use in mammalian expression.
• Viral Vectors: Engineered viruses that can deliver genetic material into host cells. Examples include retroviral vectors, lentiviral vectors, and adenoviral vectors, each with unique characteristics that make them suitable for different applications.
• BAC and YAC Vectors: Bacterial Artificial Chromosomes (BACs) and Yeast Artificial Chromosomes (YACs) are used for cloning large DNA fragments and are useful in situations where the gene of interest is too large to be accommodated by plasmid vectors.

For a detailed overview of vector types and their applications check out our blog: “Gene Integration in Antibody Production: Overcoming Challenges for Optimal Clone Selection“.

Growth Characteristics:

• Adaptability to Growth Conditions: The ability of a cell line to adapt to specific culture conditions, such as growth in suspension versus adherent cultures, and in serum-free media, is crucial for scalability. CHO cells, for example, are highly adaptable to suspension culture, which is ideal for large-scale production in bioreactors.
• Doubling Time: Faster-growing cell lines can shorten production timelines. However, a balance must be struck between growth rate and the quality of protein expression, as rapidly dividing cells may sometimes produce lower quality or improperly folded proteins.

Protein Expression Levels:

• Yield of Protein Production: The efficiency of protein production in the chosen cell line is a key consideration, especially for industrial applications. CHO cells are often preferred for their high yield of therapeutic proteins, while HEK293 cells are chosen for producing viral vectors with high transfection efficiency.
• Post-Translational Modifications: The ability of the cell line to perform necessary post-translational modifications, such as glycosylation, phosphorylation, and disulfide bond formation, is essential for the biological activity and stability of the produced proteins. For instance, CHO and NS0 cells are chosen for their human-like glycosylation patterns, which are critical for the efficacy of therapeutic proteins.

Genetic Stability:

• Long-Term Stability: The genetic stability of the cell line over multiple passages is vital to ensure consistent production of the protein or therapeutic compound over time. CHO cells are known for their genetic stability, making them reliable for long-term production.

Transfection Efficiency:

• Ease of Genetic Manipulation: The ability to efficiently transfect and genetically manipulate the cell line is critical, particularly in the early stages of stable cell line development. HEK293 cells are often used for initial expression studies due to their high transfection efficiency, while more stable lines like CHO may be used for final production stages.

Regulatory Acceptance:

• Compliance with Industry Standards: The chosen cell line should have a history of regulatory approval for use in pharmaceutical production. CHO cells, for instance, have been used in the production of numerous FDA-approved biologics, making them a safe and accepted choice for new biopharmaceuticals.

Transfection Methods for Stable Cell Line Generation

Chemical Transfection Methods

• Lipofection: Lipofection involves the use of cationic lipids that encapsulate the DNA, forming liposomes that fuse with the cell membrane to deliver the genetic material into the cell. This method is widely used due to its simplicity and effectiveness in a variety of cell types. While lipofection is primarily associated with transient transfection, it can also be used to achieve stable transfection when combined with a selection process.
• Calcium Phosphate Transfection: This traditional method involves the formation of a calcium phosphate-DNA precipitate, which is taken up by the cells. It is cost-effective and has been used extensively in the past, particularly for adherent cell lines like HEK293. This method remains a viable option for creating stable cell lines, especially in laboratories with budget constraints. However, it is less commonly used today due to the availability of more efficient and less variable methods.

Physical Transfection Methods

• Electroporation: Electroporation uses electrical pulses to create temporary pores in the cell membrane, allowing DNA to enter the cell. This method is particularly effective for difficult-to-transfect cells, such as CHO cells, which are commonly used in biopharmaceutical production. Electroporation is highly effective for generating stable cell lines, particularly for large constructs or when high transfection efficiency is required. The method’s ability to transfect both adherent and suspension cells makes it versatile and widely applicable.
• Nucleofection: This is a specialized form of electroporation that delivers DNA directly into the cell nucleus, significantly enhancing transfection efficiency. This method is particularly useful for primary cells and other cell types that are resistant to conventional transfection methods. Nucleofection is highly effective for generating stable cell lines, especially when working with hard-to-transfect cells. The direct delivery to the nucleus increases the chances of genomic integration and stable expression.

Viral Transduction

• Lentiviral Transduction: Lentiviruses are highly efficient at delivering genetic material into a wide range of cell types, including non-dividing cells. The viral genome integrates into the host cell DNA, ensuring stable and long-term expression of the transgene. Lentiviral transduction is particularly useful for generating stable cell lines when high efficiency and stable integration are required. This method is often used in both research and therapeutic applications, such as gene therapy.

Other Transfection Method

• CRISPR/Cas9-Mediated Transfection: Although traditionally used for gene editing, CRISPR-Cas9 can also be used to insert genes at specific locations in the genome by introducing double-strand breaks at specific genomic locations that are then repaired by the cell’s natural repair mechanisms, allowing the transgene to be integrated. This method is gaining popularity because its precision reduces the risk of off-target effects and ensures that the transgene is integrated at the desired genomic location.

The Complexity of Stable Cell Line Generation

Why Expertise Matters

The process of generating stable cell lines is far from straightforward – it requires a deep understanding of molecular biology, genetics, and cell culture techniques. Expertise in these areas is critical because each step, from vector design to clone selection, must be meticulously planned and executed to ensure the desired results. Experienced researchers bring invaluable insight into the nuances of cell behavior, transfection efficiency, and gene expression stability that are critical to developing robust and reliable cell lines.

At ProteoGenix, our team of experts has honed their skills through over 30 years of dedicated research and hands-on experience in cell line development, backed by over 100 successful projects. We understand the complexities involved and have developed streamlined protocols that minimize risk and maximize success. This level of expertise is not just about technical skills; it’s about knowing how to anticipate challenges and adjust strategies in real-time to ensure that the final product meets both scientific and regulatory standards.

Common Pitfalls in cell line development

Researchers can encounter several challenges during stable cell line development that may impact the success of the project:

• Vector Design Optimization: Achieving stable integration and consistent expression requires a deep understanding of promoter activity, enhancer elements, and selectable markers. Ensuring that these components are correctly chosen and configured is crucial to avoid issues with gene expression stability.
• Transfection Efficiency: The choice and optimization of the transfection method are vital for the success of stable cell line generation. Matching the transfection technique to the specific cell type is key to achieving high transfection efficiency and maintaining cell viability.
• Clone Selection: Identifying and isolating high-expressing, genetically stable clones is one of the most critical steps in cell line development. The process requires careful screening and selection to ensure that the final cell line meets the desired criteria for consistency and performance.
• Regulatory Navigation: Understanding and adhering to the regulatory requirements for cell line development is essential. Ensuring compliance with these standards from the outset helps to prevent delays and ensures that the cell line is ready for commercial or clinical use.

When to Seek Professional Services

While many research teams possess the skills and knowledge to undertake stable cell line development, there are specific situations where seeking external expertise can significantly enhance the success and efficiency of the project. Here are key indicators that it might be time to consider professional services:

• Complex Project Requirements: If your project involves complex genetic modifications, such as multi-gene insertions or precise gene editing (e.g., CRISPR/Cas9), external expertise can provide the advanced technical support needed to navigate these challenges effectively.
• Tight Timelines: When project deadlines are strict, partnering with a professional service can accelerate the development process. Experienced teams can streamline workflows, reduce trial-and-error phases, and meet critical milestones faster.
• Regulatory Compliance Needs: If your cell line is intended for therapeutic production or other applications subject to stringent regulatory oversight, professional services can ensure that all regulatory requirements are met, reducing the risk of delays or non-compliance.
• Limited In-House Resources: When your lab lacks the specialized equipment or personnel required for certain stages of cell line development, such as high-throughput clone screening or advanced transfection methods, external services can fill these gaps efficiently.
• High Failure Rates in Previous Attempts: If previous attempts at stable cell line development have resulted in inconsistent or unsatisfactory outcomes, bringing in professional services can help diagnose and resolve underlying issues, improving overall success rates.

Overview of ProteoGenix’s Custom Cell Line Development Services

ProteoGenix’s Custom Cell Line Development Service is designed to address and overcome the critical challenges associated with stable cell line generation such as optimal clone selection in antibody production.

• Early assessment of developability: We focus on an extensive preliminary testing phase, this service ensures that only the most stable, and effective cells are selected for large-scale production, minimizing risk and optimizing costs. This detailed analysis allows us to identify cells with the highest potential for stable expression and therapeutic efficacy.
• Diverse Cell Lines: We offer a wide range of non-IP cell line options, including versatile cell lines, such as our proprietary CHO-K1 and HEK293 cell lines. These cell lines are preferred for their robust expression capabilities and flexibility, allowing for optimal gene amplification and significantly higher production levels.
• Optimal clone selection: To improve the selection of optimal clones, ProteoGenix uses advanced technologies such as the Verified In-Situ Plate Seeding (VIPS) system. VIPS enables accurate and efficient single-cell cloning, ensuring the selection of highly productive and stable clones. The technology significantly reduces the time and effort associated with traditional methods.

Learn more about our custom cell line development services

Choosing ProteoGenix means leveraging our extensive expertise, innovative technologies, and commitment to quality, all of which contribute to successful antibody production projects.

Book a call with our experts today to discuss your project needs!

References

Wang, Z., Wu, X., Chen, J., Zhang, Y., & Hu, Z. (2021). Advances in the generation of stable cell lines. Frontiers in Bioengineering and Biotechnology, 9, 806791. https://doi.org/10.3389/fbioe.2021.806791