Optimizing Sample Preparation Approaches for Quantification of PROTACs

A proteolysis targeting chimera (PROTAC) is a heterobifunctional molecule composed of two active domains and a linker. One of the active domains is specific for the protein target of interest that needs to be degraded, while the other domain recruits the protein degradation machinery of the cell and brings it in close proximity to the protein target resulting in its degradation. The bioanalytical landscape for these molecules is evolving, as there may be a need to quantify the intact PROTAC's molecule, as well as the individual small molecule drugs in the same assay to maximize the information obtained from a small volume of sample.

Here, we screen multiple routinely used sample preparation techniques against a panel of PROTACs molecules spiked in biological fluids and evaluate parameters such as recoveries, matrix effects and phospholipid removal to recommend the best approach to achieve maximum sensitivity for these molecules.