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.
Techniques of Cell Line Development
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 CloneSelect Single-Cell Printer enables the gentle isolation of cells in a manner that maximizes cell viability, as well as providing direct evidence of clonality via a series of 5 images captured as cells dispense.
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.
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.
Clone Productivity Screening
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 multi-step process that involves isolating single cells from limiting dilution followed by assessment of titer using ELISA. The ClonePix 2 System combines single-cell isolation and productivity screening into a single step, resulting in dramatically shorter screening times and increased number of candidates.
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 cell surface clones 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.
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