Methods: Whole saliva from four healthy volunteers was collected and subjected to affinity based amylase removing device (provisional patent application 60915204). The filtrate was analyzed by activity assay, protein composition by SDS-PAGE, 2-DE and identified by Mass spectrometry (MS). Results were reported only when p<0.005 using t-test. The captured amylase was eluted from the device and analyzed by SDS-PAGE and MS. Results: Depleted whole saliva was analyzed by SDS-PAGE showing at least six fold removal efficiency and by an alpha amylase activity assay showing 97% reduced activity. 2-DE analysis showed the selective removal of alpha amylase and consequently increased gel resolution. MS identification of protein spots in the 60 kDa area revealed 13 spots and 15 new proteins which were masked before alpha amylase removal. Specificity was proven by elution of the captured amylase followed by SDS-PAGE and MS. Alpha amylase content was 86% from total protein content.
Conclusions: Treatment of human whole saliva with an alpha amylase removing device increases both gel resolution and enabled a higher protein sample for analysis