Cell Line Development
Stable cell lines are widely used in a number of important applications including biologics (e.g. recombinant protein and monoclonal antibody) production, drug screening, and gene functional studies. The process of developing stable cell lines often starts with transfecting selected host cells, typically CHO or HEK 293 cells, with desired plasmids. After transfection, researchers then screen and quantify high-expressing clones. Once these high producers are identified, the cell lines and/or the proteins produced by the cells are validated.
The manual screening methods traditionally used for cell line development are time-consuming and labor-intensive, creating a great demand for high throughput, automated solutions for such efforts. The general workflow below helps identify the systems that can aid in your research.

Cell line development process:
- Transfection is the process of introducing a foreign DNA (encoding the recombinant protein of interest) into a host cell. A small population of cells with the foreign DNA integrated into their genome that maintain their ability to express recombinant protein for long periods of time are referred to as stably transfected cells.
- Antibodies screening/titer ranking – Discovery and selection of high-value clones from a transfected pool of cells. To screen large populations by quantifying cell surface expression of protein-of-interest or secreting antibodies (titer ranking) will increase the probability of finding rare high-affinity binder or high producer.
- Single-cell isolution and cell viability – Single, viable cells must be isolated and cloned in order to ensure that the cell population is genetically identical, significantly reducing the heterogeneity of expression
- Monoclonality assurance – When developing cell lines for biotherapeutics, it is crucial from a quality and regulatory perspective to ensure that the cell line originates from a single progenitor and is therefore monoclonal. Documentation of monoclonality (a regulatory metric for therapeutic cell lines) is typically image-based, whereby an image of a single cell is recorded and included in regulatory filings.
- Clone productivity screening and titer – This is a test that detects the amount of recombinant protein or antibodies produced from the clonally-derived cell line.
Applications and methods for the development of cell lines
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Cell surface expression screening
Many proteins that express to the surface of cells are targets for the discovery and development of biopharmaceuticals. For instance, G-protein coupled receptors (GPCRs) are the largest class of cell-surface proteins and are targets for almost 40% of existing drugs. Discovery and selection of high value clones with elevated cell surface expression of GPCRs from a transfected pool of cells can be challenging. The ClonePix 2 System represents an automated method of screening large populations of cells that increases the probability of finding rare high-affinity binder or high producer.
Cell viability
Cell line development requires the discovery of single cell-derived clones that produce high and consistent levels of the target therapeutic protein. A critical first step in the process is the isolation of single, viable cells. Single cells proliferate to form colonies that can then be assessed for productivity of the target therapeutic protein. Viability and growth rates of single cell-derived clones can be characterized on the CloneSelect Imager and the SpectraMax i3x microplate reader and imaging cytometer.
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Clone productivity screening and titer
An important component in identifying high-value clones is determining productivity of single cell-derived colonies. Screening for productivity using traditional approaches is laborious and time consuming, generally consisting of a multistep process that involves isolating single cells from limiting dilution followed by assessment of titer using ELISA. The ClonePix 2 system combines phenotype selection, single-cell isolation and productivity screening into a single step, resulting in dramatically shorter screening times and increased number of candidates.
Colony Picking
Colony picking is an essential step in biological research as scientists often need to isolate high-value clones in order to mass-produce DNA or proteins to be used in a variety of applications downstream. Traditionally, microbial colony picking is performed manually using sterile pipette tips or inoculation loops, which are usually slow, labor-intensive, and time-consuming. Mammalian colonies are typically selected with tedious processes like limiting dilution that can negatively affect cellular outgrowth. Not only do automated colony pickers make the entire process quicker, but the results are more consistent and repeatable.
Learn more about our automated colony pickers:
- The QPix® Microbial Colony Picker leverages best-in-class colony picking technology to alleviate bottlenecks and quickly, accurately, and efficiently screen through massive genetic libraries. Learn more
- The ClonePix® 2 Mammalian Colony Picker is a fully automated system for selecting high-value clones used in antibody discovery and cell line development. Screen more clones in less time with monoclonal verification on day zero, then screen and identify for highest producers in weeks, not months. Learn more
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COVID-19 and Infectious Disease Research
Here we've addressed common applications in infectious disease research including cell line development, binding affinity, viral neutralization, viral titer and more with a focused effort on understanding the SARS-CoV-2 virus in order to develop potential therapies for COVID-19 including vaccines, therapeutics and diagnostics.
IgG Quantification
In therapeutic protein engineering and cell line development, IgG quantification is the process of measuring the amount of immunoglobulin G (IgG), a common class of therapeutic proteins, produced by a genetically modified cell line. This is important for evaluating and monitoring the productivity of the cell line and selecting the best candidate for further development of therapeutic antibodies.
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Monoclonality
Cell line development and assurance of monoclonality are critical steps in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. A cell line can be established following the isolation of a single viable cell robustly expressing the protein of interest. A key milestone in this process is documenting evidence of clonality. Documentation of clonality is typically image-based, whereby an image of a single cell is produced and included in regulatory filings.
Single-Cell Sorting
Cell line development requires the discovery of single cell-derived clones that produce high and consistent levels of the target therapeutic protein. The first step in the process is the isolation of single, viable cells. Limiting dilution is a technique that relies on statistical probability but is time consuming. The DispenCell™ Single-Cell Dispenser enables fast, easy, and gentle isolation of cells, as well as provides instantaneous proof of clonality and traceability post-cell dispensing.
Latest Resources
Resources of Cell Line Development
Brochure
Molecular Cloning
Molecular Cloning
Our solutions for antibody discovery and cell line development provide dedicated, scalable, and easy-to-use products for establishing clonal populations. The systems feature a selection of o…
Scientific Poster
Single-cell dispensing and screening of cell lines for monoclonality verification using the impedance-based single-cell dispenser and high-throughput fluorescence-based imager
Single-cell dispensing and screening of cell lines for monoclonality verification using the impedance-based single-cell dispenser and high-throughput fluorescence-based imager
There is an unmet need in the industry for a device that allows the fast and efficient isolation of single cells while preserving their integrity and providing insurance for their clonality.…
Scientific Poster
Optimizing animal-free CloneDetect assay to maximize the sensitivity of real-time local detection of human IgG antibody production using ClonePix system
Optimizing animal-free CloneDetect assay to maximize the sensitivity of real-time local detection of human IgG antibody production using ClonePix system
How to screen and select clones of interest with high specificity and high efficiency is known as one of the bottlenecks for therapeutic antibody engineering and cell line development. When…
Application Note
To achieve over 5 fold of human IgG protein detection signal enhancement with 1 round of CloneDetect K8495 assay optimization
To achieve over 5 fold of human IgG protein detection signal enhancement with 1 round of CloneDetect K8495 assay optimization
Protein G (Pro G) is a large cell surface binding protein that has a repeating 55-residue domain that binds with high affinity to the FC region of IgG. Together with Protein A (Pro A),…
Application Note
High-sensitive and affordable single-cell RNA sequencing
High-sensitive and affordable single-cell RNA sequencing
Single-cell RNA sequencing (scRNA-seq) is a state of the art technology for the unbiased characterization of gene expression in complex cell and tissue samples. Despite having…
Brochure
DispenCell™ Single-Cell Dispenser
DispenCell™ Single-Cell Dispenser
DispenCell™ is an automated laboratory instrument developed for fast, easy and gentle single-cell isolation. Designed by scientists for scientists, DispenCell has been designed to integrate…
Brochure
BioAssemblyBot 400 (BAB400): more than a bioprinter
BioAssemblyBot 400 (BAB400): more than a bioprinter
The BioAssemblyBot® 400 is an intelligent robot used by life scientists to build 3D model systems with increased throughput and precision, alleviating major concerns mentioned above.
Application Note
Automation of the CRISPR-based cell line development workflow
Automation of the CRISPR-based cell line development workflow
An increase in demands for faster high-throughput cell-line development and better screening in the pharmaceutical space is driving research facilities and companies to automate existing…
Publications
Q&A: What the transition away from animal testing could mean for drug discovery
Q&A: What the transition away from animal testing could mean for drug discovery
In September, the U.S. Senate unanimously passed the FDA Modernization Act 2.0, which would lift an 84-year-old federal mandate for animal testing for toxicity studies. While the bill doesn’…
News
Molecular Devices expands global R&D hub in Austria
Molecular Devices expands global R&D hub in Austria
Larger site will be future home of the Organoid Innovation Center – Salzburg, a collaborative space for advancing automated cell line development, organoid development, and screening solutio…
Brochure
Molecular Devices: A life sciences company
Molecular Devices: A life sciences company
Molecular Devices
Blog
[Podcast] Challenges of traditional cell line development and emerging technologies to help regulate monoclonality
[Podcast] Challenges of traditional cell line development and emerging technologies to help regulate monoclonality
Advancements in genetic engineering and synthetic biology have allowed numerous breakthroughs in recent decades. The importance of cell line development needs an honorable mention.…
eBook
How to Implement Colony Picking Workflows with Automation
How to Implement Colony Picking Workflows with Automation
Colony picking represents a powerful tool within the workflow of mammalian cell line development and the study of microbiology.
Application Note
CloneSelect Imager FL fluorescence for rapid day zero monoclonality
CloneSelect Imager FL fluorescence for rapid day zero monoclonality
Gene editing allows DNA manipulation, i.e., addition, deletion, or modification using different tools. Several nucleases for genome editing are now available: zinc finger nucleases (ZFN),…
Blog
The role of Monoclonal Antibodies against COVID-19
The role of Monoclonal Antibodies against COVID-19
Discover why mAbs are key in the fight against SARS-CoV-2 and how the pandemic has shaped their discovery and development pipeline. Over the past three years, the development of…
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COVID-19 Timeline: Diagnostics, Vaccines, and Therapeutic Antibody Development
COVID-19 Timeline: Diagnostics, Vaccines, and Therapeutic Antibody Development
Global research efforts are focused on understanding the SARS-CoV-2 virus in order to develop potential therapies for COVID-19. Join us as we explore a scientific timeline of key…
Blog
How to Develop Monoclonal Cell Lines for Stem Cell Engineering
How to Develop Monoclonal Cell Lines for Stem Cell Engineering
Monoclonal assurance is undeniably critical in cell line development for generating therapeutics, such as monoclonal antibodies or cell-based therapies. To utilize a cell line for…
Blog
Lab Automation 101: Inside Access From Our Subject Matter Expert
Lab Automation 101: Inside Access From Our Subject Matter Expert
In life sciences, more is more. Obtaining a great body of data that is consistent and insightful to get a qualitative and quantitative overview of your model is critical. However,…
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Accelerated Production of Monoclonal Antibodies for the Treatment of SARS-CoV-2
Accelerated Production of Monoclonal Antibodies for the Treatment of SARS-CoV-2
With solutions from Molecular Devices, scientists can fast-track the FDA approval process and accelerate monoclonal antibody discovery The timeline to identify and develop…
Brochure
Clone screening solutions
Clone screening solutions
Advance your microbial and mammalian clone screening with proven, automated technologies
Blog
The evolution of CHO cells’ role in cell line development
The evolution of CHO cells’ role in cell line development
Since the first approval of recombinant insulin and human growth hormone in the early 1980s, a multitude of recombinant protein therapeutics have been approved by regulatory…
Brochure
ClonePix 2 Mammalian Colony Picker
ClonePix 2 Mammalian Colony Picker
Redefine clone screening and selection with transformative cell line development workflows.
Flyer
High-throughput stem cell screening
High-throughput stem cell screening
Regenerative medicine has the potential to provide cures for some of the world’s most devastating diseases. However, many roadblocks remain when it comes to moving these therapies from the b…
Blog
Get to know our Field Applications Scientist: Dwayne Carter
Get to know our Field Applications Scientist: Dwayne Carter
Dwayne Carter gives us a taste of 3D bioprinting, clone screening, and Caribbean cuisine Dwayne Carter is a cell biologist and educator who joined Molecular Devices in November 2020…
Publications
Advanced Cell Screening and Imaging Reveals How the Cardiac System Responds in Drug Development
Advanced Cell Screening and Imaging Reveals How the Cardiac System Responds in Drug Development
In drug discovery, time and monetary investments increase significantly the further candidates advance through the preclinical phase of testing, analysis and evaluation. By excluding non-via…
Application Note
Measuring cell viability and cytotoxicity with the EarlyTox Cell Integrity Kit
Measuring cell viability and cytotoxicity with the EarlyTox Cell Integrity Kit
The EarlyTox™ Cell Integrity Kit is an optimized set of reagents that simplifies the identification of live and dead cells. It can be used to measure the effects of different treatments on…
Application Note
A microfluidics-based, single cell printing and microplate imaging workflow optimized for efficiency, viability and monoclonality report generation
A microfluidics-based, single cell printing and microplate imaging workflow optimized for efficiency, viability and monoclonality report generation
In this app note, we demonstrate a unified workflow that combines single cell deposition and clonality verification. We couple the use of the SCP and CSI to deposit single cells into a…
Application Note
A workflow combining single-cell isolation and microplate imaging improves cloning efficiency with higher assurance of clonality
A workflow combining single-cell isolation and microplate imaging improves cloning efficiency with higher assurance of clonality
The isolation of single cells has a broad spectrum of applications, from single-cell genomics to antibody discovery and cell line development.
Application Note
Confident assurance of clonality using calcein AM with minimal effect on viability
Confident assurance of clonality using calcein AM with minimal effect on viability
The development of cell lines that express a specified protein of interest is critical to the generation of biopharmaceutical molecules such as monoclonal antibodies. To establish a cell…
Scientific Poster
Rapid Screening and Selection of Stable High Producing Clones
Rapid Screening and Selection of Stable High Producing Clones
Screening cell populations for stable, high producing clones is a key rate-limiting step in the development and production of new protein therapeutics.
Brochure
CloneSelect Imager Monoclonality Report Feature
CloneSelect Imager Monoclonality Report Feature
Providing image evidence of monoclonality in the cell line development process is not as simple as exporting an image of a single cell. For example, high-resolution images of the entire well…
Scientific Poster
Rapid Monoclonality Verification Methods to Boost Cell Line Development
Rapid Monoclonality Verification Methods to Boost Cell Line Development
Here we present a fluorescent method for identifying monoclonal CHO-S cells using CellTracker Green CMDFA.
Application Note
Rapid and efficient selection of high producing mammalian cells secreting non-mAb proteins
Rapid and efficient selection of high producing mammalian cells secreting non-mAb proteins
The technology in ClonePix Systems removes the need for serial dilution or cell-sorting techniques. It allows monoclonal populations of high-secreting cells to be established in a much…
Application Note
Automated Optimization of IgG Production in CHO Cells
Automated Optimization of IgG Production in CHO Cells
Monoclonal antibodies have long been an essential tool for cell and molecular assays and have moved into the clinic as part of a shift from small molecules into biologic treatments for…
Application Note
Confident Identification of Monoclonal CHO-S Cells Grown in Semi-Solid Media using the CloneSelect Imager
Confident Identification of Monoclonal CHO-S Cells Grown in Semi-Solid Media using the CloneSelect Imager
Regulatory criteria to obtain approval of biological drugs are constantly evolving. One of the regulatory metrics for biological cell lines, the digital documentation of monoclonality, has…
Application Note
Confluence and growth curve generation on the CloneSelect Imager
Confluence and growth curve generation on the CloneSelect Imager
The ability to monitor cell proliferation has become increasingly important, particularly with the widespread use of in vitro cell lines as model systems. Proliferation rates can be used to…
Application Note
Rapid automated selection of mammalian cell colonies by cell surface protein expression
Rapid automated selection of mammalian cell colonies by cell surface protein expression
The ability to select cells based on surface protein expression is of great use in establishing cell lines expressing the receptor(s) of interest for downstream phenotyping/genotyping,…
Application Note
Rapid selection and development of GPCR expressing mammalian cell lines using novel ClonePix Technology
Rapid selection and development of GPCR expressing mammalian cell lines using novel ClonePix Technology
Discovery and selection of high expressing GPCR clones from a transfected pool of cells can be challenging with regards to cell-line development. The technology in ClonePix 2 Systems…
Application Note
Enhanced development of virus-specific hybridomas using ClonePix and CloneSelect Imager Technologies
Enhanced development of virus-specific hybridomas using ClonePix and CloneSelect Imager Technologies
The ClonePix System was employed in the high-throughput screening of hundreds of sub-cloned colonies from parental hybridoma material (historically parental line yields were less than 1 mg/…
Application Note
Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
The CellTiter-Glo assay from Promega uses luciferase enzyme, which requires ATP in order to generate light. The luminescent signal produced in the assay depends on the amount of ATP in the…
Brochure
SpectraMax i3x Multi-Mode Detection Platform
SpectraMax i3x Multi-Mode Detection Platform
The SpectraMax i3x Multi-mode microplate reader measures spectral-based Absorbance, Fluorescence, and Luminescence with added functaionlity of modular upgrades for Western Blot, Imaging, and…
Videos & Webinars

Selection of high producing mammalian cells secreting non-mAb proteins workflow - SynBioBeta Lightning Talk

The many and varied workflows of cell line development