Establishing Personalized Blood Protein Reference Ranges Using Noninvasive Microsampling and Targeted Proteomics: Implications for Antidoping Strategies

To prevent doping
practices in sports, the World Anti-Doping Agency
implemented the Athlete Biological Passport (ABP) program, monitoring
biological variables over time to indirectly reveal the effects of
doping rather than detect the doping substance or the method itself.
In the context of this program, a highly multiplexed mass spectrometry-based
proteomics assay for 319 peptides corresponding to 250 proteins was
developed, including proteins associated with blood-doping practices.
“Baseline” expression profiles of these potential biomarkers
in capillary blood (dried blood spots (DBS)) were established using
multiple reaction monitoring (MRM). Combining DBS microsampling with
highly multiplexed MRM assays is the best-suited technology to enhance
the effectiveness of the ABP program, as it represents a cost-effective
and robust alternative analytical method with high specificity and
selectivity of targets in the attomole range. DBS data were collected
from 10 healthy athlete volunteers over a period of 140 days (28 time
points per participant). These comprehensive findings provide a personalized
targeted blood proteome “fingerprint” showcasing that
the targeted proteome is unique to an individual and likely comparable
to a DNA fingerprint. The results can serve as a baseline for future
studies investigating doping-related perturbations.