Targeted protein degradation, especially via PROTACs, offers promising approaches for addressing previously undruggable disease-associated proteins. However, clinical success hinges on factors like target selectivity, with conventional 2D cell culture models frequently failing to capture the biological complexity necessary to accurately predict in vivo efficacy and clinical outcomes. 3D cell culture models, such as FluoSphera's color-coded spheroids, which use a cell encapsulation technique, offer a more physiologically-relevant alternative for assessment of drug selectivity, efficacy, and mechanisms of action.

In our study we combine FluoSphera’s encapsulated spheroids and advanced multiplexing technology with Revvity’s high-content imaging (HCI) and no-wash immunoassays to create a robust approach to evaluation of PROTAC compounds' efficacy and selectivity in co-culture 3D format.

In our application note we describe how to perform:

• evaluation of PROTACs efficacy and selectivity in co-cultured encapsulated 3D spheroid models.
• high-resolution, quantitative analysis of cellular morphology and molecular events at the single-cell level using HCI
• highly sensitive detection of protein expression at the whole cell-level with no-wash HTRF assays.

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