Quantifying in vitro Complement-Dependent Cytotoxicity (CDC) using Advanced Flow Cytometry

Authors: Kirsty McBain, Scientist, Analytics, Sartorius | Last updated: August 10, 2023 


In recent decades, rapid progression of monoclonal antibody (mAb) development has culminated in approvals for more than one hundred therapeutics in the United States. 

mAbs enhance the immune response through three key Fc-mediated functions: antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and complement-dependent cytotoxicity (CDC). 

CDC is initiated when a mAb triggers activation of the complement cascade, which ultimately results in tumor cell lysis.

This poster video presents a simple, high-throughput, miniaturized assay for measuring CDC, using the iQue® Advanced Flow Cytometry Platform, with its integrated iQue Forecyt® software.  Thus streamlining antibody characterization and drug discovery processes.

  • Document type: Video
  • Watch time: 19 minutes

Key Takeaways

  • The iQue® Advanced Flow Cytometry Platform, together with mix and read reagents, and easy-to-follow protocols provides a simple, high-throughput solution for quantification of monoclonal antibody (mAb) complement-dependent cytotoxicity (CDC) activity.
  • CDC activity was found to be dependent on factors such as antibody isotype and target cell expression of both the target antigen and complement regulatory receptors.
  • An Incucyte® Live-Cell Analysis System may also be used to monitor the progression of CDC activity over time.
  • These workflows deliver quick and reliable quantification of mAb Fc function, with the potential to enhance antibody discovery workflows.

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