Combined approach of selective and accelerated cloning for microfluidic chip-based system increases clone specific...

Area of Science:

• Biotechnology
• Cell Line Development
• Bioprocessing

Background:

• Current cell line development (CLD) workflows aim to increase cell-specific productivity or reduce timelines.
• Existing fast-sorting methods using low-viability pools shorten CLD timelines but reduce clone recovery.
• A need exists for CLD protocols that enhance both productivity and speed without compromising clone yield.

Purpose of the Study:

• To combine early cloning with low-viability pools and IgG membrane staining on the Beacon platform.
• To concurrently increase specific productivity and shorten timelines for generating Chinese hamster ovary (CHO) clones.
• To overcome the low recovery issue associated with previous fast-sorting approaches.

Main Methods:

• Utilized the Beacon platform for cell line development.
• Implemented a combined approach of fast-sorting with low-viability pools and IgG membrane staining.
• Employed fluorescent staining to identify and enrich high-secreting cells prior to cloning.

Main Results:

• Achieved correct cell recovery, enabling full utilization of Beacon screening capacities.
• Significantly increased the specific productivity of selected clones.
• Demonstrated a synergistic effect of the combined protocols, enhancing throughput and detection of super-productive clones.

Conclusions:

• The combined protocol effectively discards non-producing and dead cells before cloning.
• This integrated approach accelerates the generation of highly productive CHO clones.
• The method optimizes the Beacon platform's power for efficient cell line selection.