Dried blood spots offer many advantages as a sample format including ease and 
safety of transport and handling. To date, the majority of mass spectrometry 
analyses of dried blood spots have focused on small molecules or hemoglobin. 
However, dried blood spots are a potentially rich source of protein biomarkers, 
an area that has been overlooked. To address this issue, we have applied an 
untargeted bottom-up proteomics approach to the analysis of dried blood spots. 
We present an automated and integrated method for extraction of endogenous 
proteins from the surface of dried blood spots and sample preparation via 
trypsin digestion by use of the Advion Biosciences Triversa Nanomate robotic 
platform. Liquid chromatography tandem mass spectrometry of the resulting 
digests enabled identification of 120 proteins from a single dried blood spot. 
The proteins identified cross a concentration range of four orders of magnitude. 
The method is evaluated and the results discussed in terms of the proteins 
identified and their potential use as biomarkers in screening programs.