A Bench-Top Approach for Isolation of Single Antibody Producing Chinese Hamster Ovary (CHO) Cells Using a Microwell-Based Microfluidic Device

Abstract

Genetically-modified monoclonal cell lines are currently used for monoclonal antibody (mAbs) production and drug development. The isolation of single transformed cells is the main hindrance in the generation of monoclonal lines. Although the conventional limiting dilution method is time-consuming, laborious, and skill-intensive, high-end approaches such as fluorescence-activated cell sorting (FACS) are less accessible to general laboratories. Here, we report a bench-top approach for isolating single Chinese hamster ovary (CHO) cells using an adapted version of a simple microwell-based microfluidic (MBM) device previously reported by our group. After loading the cell suspension to the device, the electrostatically trapped cells can be viewed under a microscope and transferred using a micropipette for further clone establishment. Compared to the conventional method, the invented approach provided a 4.7-fold increase in the number of single cells isolated per round of cell loading and demonstrated a 1.9-fold decrease in total performing time. Additionally, the percentage of correct single-cell identifications was significantly improved, especially in novice testers, suggesting a reduced skill barrier in performing the task. This novel approach could serve as a simple, affordable, efficient, and less skill-intensive alternative to the conventional single-cell isolation for monoclonal cell line establishment.

Keywords

• monoclonal cell lines
• cell line generation
• single-cell cloning
• limiting dilution
• microfluidic device