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Cell Counting and Image Cytometry

High-Throughput Clone Screening Ensures High-Quality Biologic Production

Section
Cell-based Assays for Bioprocessing
Celigo Applications
Cell Counting Method Selection
Cell Counting and Image Cytometry FAQs
Cell-based Assays for Bioprocessing
Cell-based Assays for Gene Therapy Development
Cellometer Applications
Modern Virology Assays
Sub Section
Bioprocessing Solutions
Bioprocessing Solutions
FDA Regulations of Biologics
Topic
High-Throughput Clone Screening Ensures High-Quality Biologic Production
Engineering CHO Cells Using CRISPR Technology
High-Throughput Clone Screening Ensures High-Quality Biologic Production
Media Selection for Optimization of Culturing Conditions
Monitoring Growth and Viability
Using Cellaca MX for High-Throughput CHO Cell Counting for Bioprocessing
Single Cell Cloning for Cell Line Development

Not all problems are solved once a single clone is selected. While CHO cells are an excellent production system, there are two major issues that hamper biologic productivity [Dahodwala & Lee, 2019]:

  • Chromosomal rearrangements and stability issues
  • A phenotype characterized by titer instability and product quality instability

Following SCC, constant monitoring is needed to ensure that the clonal pool is table and producing a consistent product. Cultured cells can show decreased recombinant protein production over many passages and during extended periods at the final bioreactor scale [Tharmalingam et al. 2018].

Revvity can be used to monitor cell populations produced from a clonally derived clone, including assessment of:

  • Performance
  • Productivity
  • Product quality characteristics

In collaboration between Revvity and Beth Israel Deaconess Medical Center, Zhang et al. [2017] employed the Celigo™ image cytometer to screen for high protein producers from hybridoma supernatants. Fluorescence labeling of CHO cells (left) with a CD39 antibody was used to determine the amounts of target protein they produced. Different levels of target protein binding showed various fluorescence intensities, and the high intensities were identified as possible hybridoma candidates (green wells, right). The results were validated with standard flow cytometry.

img-high-throughput-clone-screening-ensures-high-quality-biologic-production-1.PNG

Fluorescence labeling of CHO cells with CD39 antibody

img-high-throughput-clone-screening-ensures-high-quality-biologic-production-2.PNG

High-intensity levels of target protein binding indicated by green wells

For research use only. Not for use in diagnostic procedures.

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