Key highlights:
- CD16-dependent NK cell engagement quantified under physiological flow
- Donor-specific variation in Cetuximab-driven avidity revealed
- Mechanistic insight beyond conventional cytotoxicity assays
- Paired avidity and functional data enabling confident cell population selection
The challenge
NK:IO is developing off-the-shelf allogeneic natural killer (NK) cell therapies to enable scalable, cost-effective immunotherapy. As part of their development programme, the team is investigating cell engager strategies to enhance NK cell efficacy against tumour targets, including the use of Cetuximab, a therapeutic antibody that redirects NK cells to EGFR-expressing cancer cells via CD16-mediated antibody-dependent cellular cytotoxicity (ADCC).
During preclinical development, NK:IO encountered inconsistent results from conventional cytotoxicity assays when comparing CD16⁺ and CD16⁻ NK cell populations in the presence of Cetuximab. These conflicting findings raised questions about data reliability and created uncertainty around which NK cell population would provide optimal therapeutic performance. Standard endpoint assays were unable to provide the mechanistic insight needed to make informed decisions about cell population selection.
Our Approach
We applied our avidity-based microfluidic platform to directly measure NK cell engagement with tumour targets under controlled shear forces designed to reflect physiological flow conditions, enabling a direct, side-by-side comparison of CD16⁺ and CD16⁻ NK cell populations under Cetuximab-mediated conditions, alongside parallel functional validation through matched cytotoxicity assays.
- Quantified total binding strength under controlled flow conditions
- Captured both CD16-mediated and native receptor interactions
- Enabled direct comparison of different NK cell populations
- Revealed mechanism of action insights beyond endpoint measurements
maintain cell phenotype
to final data delivery
Our Approach
We applied our avidity-based microfluidic platform to directly measure NK cell engagement with tumour targets under controlled shear forces designed to reflect physiological flow conditions, enabling a direct, side-by-side comparison of CD16⁺ and CD16⁻ NK cell populations under Cetuximab-mediated conditions, alongside parallel functional validation through matched cytotoxicity assays.
- Quantified total binding strength under controlled flow conditions
- Captured both CD16-mediated and native receptor interactions
- Enabled direct comparison of different NK cell populations
- Revealed mechanism of action insights beyond endpoint measurements
maintain cell phenotype
to final data delivery
maintain cell phenotype
to final data delivery
Key Outcomes
By assessing Cetuximab-mediated NK–target binding in a dynamic, cell-based context, we identified which NK cell population is best suited for use with Cetuximab. We also demonstrated that functional assays alone cannot reliably predict in vivo performance — insight that standard binding or cytotoxicity assays cannot provide.
Avigen was highly professional and a pleasure to work with, delivering high-quality data on a rapid timeline. Their avidity-based analysis provided valuable insight beyond standard assays and provided a new layer of validation data for future decision-making.


