Inactivation of West Nile virus in serum with heat, ionic detergent, and reducing agent for proteomic applications

•WNV in serum can be rendered non-infectious using SDS-PAGE sample buffer and heat.•This method effected a reduction in infectivity of ≥6.5×103 (≥99.98%).•This method enables proteomic analysis of samples outside of BSL-3 containment.•Proteomics of WNV-infected mouse sera identified differentially expressed proteins. Research involving biosafety level 3 pathogens such as West Nile virus (WNV) is often limited by the limited space and technical constraints of these environments. To conduct complex analytical studies outside of high containment, robust and reliable inactivation methods are needed that maintain compatibility with downstream assays. Here we report the inactivation of WNV in spiked serum samples using a commercially available SDS-PAGE sample buffer for proteomic studies. Using this method, we demonstrate its utility by identification proteins differentially expressed in the serum of mice experimentally infected with WNV.