Application Note: Evaluating ADCs In Vitro Using 3D Tumor Spheroid Models
Authors: Kirsty McBain, Kalpana Barnes & Nicola Bevan | Last updated: February 2024
Overview
Antibody-drug conjugates (ADCs) are transforming the landscape of cancer therapy by marrying the precision of targeted drug delivery with the potency of chemotherapeutic agents, all while engaging the body's own immune system to fight cancer cells. As ADC development surges forward, the demand for sophisticated and dependable assessment methods for these promising drugs has never been greater.
The use of 3D advanced cell models can improve the translational ability of these in vitro techniques. Incucyte® Live-Cell Analysis data demonstrated greater induction of cytotoxicity by anti-HER2 ADCs (Kadcyla® and Enhertu®) compared to the monoclonal antibody (mAb) backbone on which they were based (Trastuzumab).
iQue® HTS analysis revealed the unique Enhertu® bystander activity, by allowing the cellular composition of a spheroid co-culture to be measured. These techniques facilitate in-depth analysis of ADC activity and allow mechanistic differences between ADCs to be unpicked.
- Document type: Application Note
- Page count: 10
- Read time: 10 minutes
Workflow Advantages
- 3D single and multi-spheroid models enable greater insights
- Live-cell analysis provides visual and temporal analysis of spheroids over time
- High throughput screening by cytometry facilitates high-throughput analysis of dissociated cells
Figure 2. Internalization of ADCs into BT474 spheroids is greater than internalization of Trastuzumab. BT474 cells (4 K/well) were seeded into ultra-low attachment (ULA) plates for 72 hours to promote spheroid formation. Antibodies were labeled with Incucyte® Human Fabfluor-pH Orange Antibody Labeling Dye then added to spheroids (n=3). Phase and fluorescence images (10X objective) were captured every 15 minutes using the Incucyte® Live-Cell Analysis System. (A) Representative Incucyte® images of single spheroids taken 48 hours after antibody addition. (B) Internalization was quantified as an increase in Spheroid Orange Mean Intensity (OCU) and plotted over time.
