Analysis of multiple proteins is possible from one DBS sample by multiplexed MRM. Urea/Thiourea digestion resulted in the highest number of observable proteins from DBS analysis with 60percent observed. NaDOC and Urea digests also showed a comparable level of observable proteins to Urea/Thiourea, with 53percent observed. There are no concerns of peptides being introduced as a result of digestion of the filter paper itself. Reproducibility experiments on Urea/Thiourea, NaDOC, and Urea digests will allow for determination of the best method for clinical DBS protein analysis. Additional investigations can also investigate the effect of filter paper materials, or the possibility of DBS trypsin digestion directly on the filter paper spot.
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