Implications of the Coffee-Ring Effect on Virus Infectivity

The factors contributing to the survival of enveloped viruses (e.g., influenza and SARS-CoV-2) on fomite surfaces are of societal interest. The bacteriophage Phi6 is an enveloped viral surrogate commonly used to study viability. To investigate how viability changes during the evaporation of droplets on polypropylene, we conducted experiments using a fixed initial Phi6 concentration while systematically varying the culture concentration and composition (by amendment with 2% fetal bovine serum (FBS), 0.08 wt % BSA, or 0.5 wt % SDS). The results were consistent with the well-founded relative humidity (RH) effect on virus viability; however, the measured viability change was greater than that previously reported for droplets containing either inorganic salts or proteins alone, and the protein effects diverged in 1× Dulbecco’s modified Eagle’s medium (DMEM). We attribute this discrepancy to changes in virus distribution during droplet evaporation that arise due to the variable solute drying patterns (i.e., the “coffee-ring” effect) that are a function of the droplet biochemical composition. To test this hypothesis, we used surface-enhanced Raman spectroscopy (SERS) imaging and three types of gold nanoparticles (pH nanoprobe, positively charged (AuNPs(+)), and negatively charged (AuNPs(−))) as physical surrogates for Phi6 and determined that lower DMEM concentrations, as well as lower protein concentrations, suppressed the coffee-ring effect. This result was observed irrespective of particle surface charge. The trends in the coffee-ring effect correlate well with the measured changes in virus infectivity. The correlation suggests that conditions resulting in more concentrated coffee rings provide protective effects against inactivation when viruses and proteins aggregate.