TRAIL (tumor necrosis factor-related apoptosis-inducing ligand also known as APO2L - TNF superfamily) is the first recombinant human protein that selectively induces apoptosis in cancer cells while sparing normal cells. In addition to its potential anti-cancer properties, TRAIL also demonstrates numerous other potential therapeutic uses in cardiovascular diseases, diabetes and atherosclerosis. These developments make TRAIL a promising target in the field of biotherapeutics.
In developing a method, multiple challenges have to be overcome including chromatographic separation, low sensitivity and high signal variability. High Resolution Mass Spectrometry (HRMS) offers an alternative platform in quantifying TRAIL to support pharmacokinetic investigations.
To obtain successful chromatographic separation, Algorithme Pharma used a polymer based column which improved the peak shape and injection reproducibility. To increase the column recovery, the temperature was raised which resulted in a quadrupled recovery rate. The data processing method was then optimized to avoid interferences from endogenous compounds. To overcome the low sensitivity and high signal variability, the 6 most abundant multiple charge states were summed with an XIC window of 10mDa around the most abundant isotopomer.
Algorithme Pharma has successfully overcome the multiple challenges involved in using HRMS to quantify intact TRAIL and has confirmed its applicability as an optimal platform for intact protein quantification without the need for tryptic digestion.