Native Label‐Free Spectral Biochemical Fingerprinting of an Enveloped Mammalian RNA Virus Reveals Mode of Action of a Virucidal Drug
Denis Rajnovic, Fatima Matroodi, Lamyaa M. Almehmadi, Tea Carletti, Alessandro Gessini, Igor K. Lednev, Claudio Masciovecchio, Alessandro Marcello, Barbara RossiABSTRACT
Pandemic preparedness demands the availability of state‐of‐the‐art tools for fundamental research on viruses to support rapid development of antiviral and prophylactic interventions. Virion composition is currently studied by a limited number of disruptive techniques, while here, a novel workflow for non‐invasive and label‐free biochemical fingerprinting is presented. Deep‐Ultraviolet Resonance Raman spectroscopy is employed as the primary analytical tool, while stoichiometric‐based spectral reconstruction and multilinear regression analysis are used to perform bottom‐up and top‐down biochemical characterization of the vesicular stomatitis virus (VSV) in physiological‐like conditions, respectively. Raman spectra from virions and single biochemical components (lipids, amino acids and nucleotides) excited at 213 nm are used to disentangle the virus biochemical composition and to probe the mechanism of action of a lipid‐disrupting virucidal drug. Available biochemical and sequence information together with single‐component spectra allows the reconstruction of the full virion spectrum with excellent accuracy. Altogether, this strategy of virion biochemical fingerprinting paves the way to a better understanding of virus composition and mechanisms for inactivation.