Frozen whole blood cell pellets from NSCLC patients and healthy controls were processed using novel techniques to produce a unique sample using VAMS devices. Samples were analysed using liquid chromatography mass spectrometry to collectively screen 3914 proteins from the whole blood sample. Analysis of all fractions produced a set of 515 differentially expressed proteins. Boosted regression tree analysis of the differentially expressed proteins produced a panel of 13 proteins that were able to discriminate between controls and NSCLC patients with an area under the ROC curve (AUC) of 0.864 for the set. Our rapid and reproducible blood preparation and analysis methods enabled the production of high-quality data from small volumes of complex samples that are typically require significant fractionation prior to proteomic analysis.