Conquering sensitive CYPs, UGTs and transporters protein quantification with QTAP analysis

With its high sensitivity, QTAP (Quantitative targeted absolute proteomics) is also able to quantify the major CYPs, UGTs and transporters of interest.

Dr. Mouhssin Oufir will explore with you the QTAP analysis  and its application for the transporter quantitation in the context of a validation of a human in vitro Blood Brain Barrier (BBB) model.

RT-PCR and Western blot were not able to detect the two following ABC transporters, hMDR1 (P-gp) and hBCRP, involved in active transport in the hBMEC (human Brain microvascular endothelial cell). Whereas QTAP approach was able to detect but not quantifiy P-gp and BCRP in the cell membrane preparation.

The DDI (Drug-Drug interactions) guidelines (FDA and EMA) both cite these two transporter proteins as being highly relevant in ADME studies. Thus, our results reveal a major limitation for the use of the hBMEC cell line as in vitro human BBB model for the screening of drug candidates.

Key learning objectives:

• Introduction to Quantitative Targeted Absolute Proteomics
• Application of QTAP in research
• Advantages of QTAP versus Western blot or other approaches…
• Case study