Revolutionizing Cell Line Development With Cutting-Edge Automation Solutions

Danaher Corporation

In today’s complex and rapid biopharmaceutical landscape, challenges in early drug development have required key advancements in the technologies and processes to support these workflows. While many of the cell line development (CLD) processes performed today are still highly manual, there exists significant opportunity to automate discrete process steps, or to pursue more intensive degrees of laboratory automation.

Automation can take many different forms in a biopharmaceutical workflow, and is often focused on reducing the hands-on time necessary for processing a given step. To perform a cell line stability study, for example, an organization may enable a point solution such as automated sample processing by integrating a liquid handler with an incubator, eliminating the need to split cultures manually every three or four days. In other cases, organizations may work to pursue “total lab automation,” establishing a workflow where a process is managed by an interconnected system, automating processes ranging from cell culture to analytical sample preparation. While this level of automation still requires hands-on work for certain process steps, it serves to increase reproducibility and consistency, which in turn can increase laboratory capacity.

The diversity of the workflows established by developers and laboratories over the years has led to a proliferation in technologies designed to support CLD campaigns. This variability in both equipment and supporting protocols across drug development can make scaling processes and tech transfer to an external partner a challenge. Yet, advancements made in support of these workflows have created the potential for greater standardization and automation, eliminating the most common workflow bottlenecks and improving the reproducibility of a process.

Targeted Automation to Streamline Crucial CLD Workflows

Targeted Automation to Streamline Crucial CLD Workflows

Even among more thoroughly characterized modalities like monoclonal antibodies (mAbs), the uniqueness of an individual asset can require tailored approaches that balance reproducibility and standardization with the need for hands-on work. Outdated technologies and disparate instruments making up a workflow can hamper efforts to implement laboratory automation, as can outdated methods ill-equipped for automation and integration. Even so, automation can be incorporated into the majority of the process steps that typify a mAbs workflow, from stable transfection and monoclonality verification to clone selection and antibody characterization. Creating "islands of automation" can serve to streamline some of the most crucial process steps in a CLD workflow; moreover, connecting workflow steps through automation, supported by digital solutions that enable interoperability, may be possible for certain applications, and result in a more closed process with less hands-on work.

Today, there are a number of crucial CLD workflow steps that can benefit from automation solutions. While some CLD workflow steps still require more manual approaches or are more easily outsourced, others are ripe for automation. Danaher Life Sciences, through its portfolio of companies spanning the advanced therapy development and manufacturing space, has amassed a range of solutions geared toward CLD automation for the process steps that can benefit most:

Stable Transfection and Clone Selection

No matter what method a developer uses to stably transfect during cell line development, from electroporation to chemical gene delivery methods and beyond, eliminating hands-on time can go a long way toward improving consistency. Incorporating liquid-handling systems like the- Series Automated Liquid Handler can free up operators to perform other, less easily automated tasks; automating this step can hugely reduce the potential for error that is nearly unavoidable when performing thousands oftransfectionsby hand. By delegating this work to a liquid handler, personnel can focus on more intensive tasks.

Once cell lines have been generated, they are screened for key critical quality attributes (CQAs), and inoculated into culture.. While these steps are often performed independently, all-in-one technology solutions like the Molecular Devices ClonePix 2 Mammalian Colony Picker can allow for clone screening and culture in a single instrument solution. Automating clone selection is a valuable undertaking, as it affords workflows greater precision than can typically be achieved through manual selection. Technologies like the ClonePix 2 Mammalian Colony Picker can are out-of-the-box automation solutions for more than one step of this workflow, bringing greater value to organizations performing extensive clone selection.

Once transfected cell lines have been selected for culture, samples are taken from each culture to measure viability. This step is often performed using a trypan blue dye exclusion test, with operators manually counting and determining cell viability from images containing thousands of cells. This step is well-positioned for automation using tools like the Beckman Coulter Life Sciences Vi-CELL BLU Cell Viability Analyzer, which can perform completely automated sample analysis in seconds.

Monoclonality Verification

Confirming that cell cultures have originated from a single cell is a complex, time-consuming part of cell line development. Operators must be able to demonstrate monoclonality with time series data, and failing to demonstrate monoclonality can prevent a promising cell line from entering the master cell bank (MCB). Instruments like the Biomek i-Series Automated Liquid Handler can help to demonstrate monoclonality, and cell sorters like Beckman Coulter Life Sciences’ CytoFLEX SRT Benchtop Cell Sorter can employ complex sorting logic, perform independent routine maintenance, and automate a sorting stream, as well as enable complex sort logic across four sorting streams. This flexibility, coupled with a small laboratory footprint and simplified setup and operation, can serve to reduce downtime and enable crucial insights without the need for complex operational expertise.

Another fit-for-purpose tool to support monoclonality verification is Molecular Devices’ Clone Select FL Imager, which offers time-series imaging of colony formation from a single cell, with a number of key features, including:

More fit-for-purpose technologies like the ClonePix 2 Mammalian Colony Picker can offer operators compounding benefits – on top of performing clone selection and cell culture, the ClonePix 2 Mammalian Colony Picker can also verify monoclonality on day zero using high-resolution, single-cell imaging with white light and fluorescence detection. Furthermore, solutions such as Molecular Devices’ DispenCell Single-Cell Dispenser can offer organizations fast, easy cell isolation supported by real-time analysis software that allows users to validate clonality immediately after a cell is dispensed.

Selection Based on Stability and Expansion

Cell line expansion, which involves serially passaging cells from smaller cultures to larger ones, is the CLD step that enables operators to quantify a line’s productivity and long-term stability. Stability studies are both a critical assessment of a cell line’s viability and a heavy lift for most organizations – characteristics such as product titer, product CQAs, and clone robustness must be measured to generate insights that can inform downstream processes.

Automating the analytical approaches at the core of this paradigm can help operators achieve more streamlined workflows supported by in-house data generation. Solutions like Beckman Coulter Life Sciencess Valita Titer Assays for IgG Quantification can enable rapid, cost-effective, automation-friendly IgG measurement using cell suspension, and can be read with any plate reader that features a fluorescence polarization module. With results in less than 15 minutes generating data of comparable quality to that of an HPLC system, Valita Titer assays can be automated using the Biomek i-Series Automated Liquid Handler in conjunction with the Molecular Devices SpectraMax microplate reader. In contrast to a traditional Enzyme-Linked Immunosorbent Assay (ELISA), the Valita Titer assays do not require sample preparation or wash steps, enabling rapid time-to-data. Alongside these assays, solutions like the Vi-CELL BLU Cell Viability Analyzer can likewise offer rapid insights on cell counts, informing operators on when to passage cultures.

Antibody Characterization

Many of the analytical evaluations that generate detailed data on antibody CQAs may be outsourced to third-party labs, owing to their complexity and the expertise and laboratory footprint necessary to bring them in-house to a CLD lab. Yet there are a number of technology solutions today that can be incorporated into a CLD workflow relatively easily while affording operators more immediate insights. Solutions like the SCIEX BioPhase 8800 system, a multi-capillary electrophoresis system, can allow operators to run up to eight samples in parallel, enabling increased capacity and throughput.

One of the biggest challenges that accompanies analytical methods such as mass spectrometry systems is sample preparation. Often, the intensive preparation needed to clean a sample to the degree necessary to introduce it to a highly sensitive mass spectrometer prompts many organizations to outsource assays that require mass analyses. Products like the SCIEX Intabio ZT system couple automated icIEF sample preparation with high resolution mass spectrometry on a SCIEX ZenoTOF 7600 System, allowing operators to access more insightful data using instruments that are that is easier to use and can be fully automated from acquisition to processing.

Conclusion

Ultimately, CLD workflows are highly complex, with many delicate, time-sensitive, and repetitive steps that require significant personnel and training to support. Many of these steps are challenging and leave little room for error or variability. Automating parts of a CLD process can radically improve the consistency and reproducibility of certain steps, which can, in turn, benefit later process development. As automation solutions rapidly increase in the biopharmaceutical industry, incorporating technologies that can offer a process more streamlining and consistency will be key to remaining competitive in an evolving market.