Amylase Depletion Significantly Improves the Maldi-Tof Profile of Saliva – Application to Covid-19 Diagnostics

Abstract

Human saliva contains a plethora of proteins whose presence and concentration can be monitored for diagnosis and progression of disease. Saliva has been extensively probed for the diagnosis of several systemic and infectious diseases because of the ease with which it can be collected. However, amylase, the most abundant protein found in saliva can obscure the detection of low-abundance proteins by MALDI-ToF MS (matrix-assisted laser desorption/ionization-time of flight mass spectrometry) and diminish the diagnostic utility of this specimen type. In the present study, we used a device to deplete salivary amylase from water-gargle samples through affinity adsorption. After depletion, profiling of the saliva proteome was performed by MALDI-ToF MS on gargle samples from subjects whose COVID-19 (coronavirus disease 2019) status was confirmed by NP (nasopharyngeal) swab RT-qPCR (reverse transcription polymerase chain reaction). Amylase depletion led to the enhancement of signal intensities of various peaks as well as the detection of previously unobserved peaks in the MALDI-ToF spectra. The overall specificity and sensitivity after amylase depletion was 100% and 85.17% respectively for detecting COVID-19. Our simple, rapid and inexpensive technique to deplete salivary amylase can be used to unmask spectral diversity in saliva by MALDI-ToF MS, reveal low-abundant proteins and aid in the establishment of novel biomarkers for diseases.