A gradient-free method for the purification of infective dengue virus for protein-level investigations

•A 5-h protocol to purify dengue virus from contaminants, including C6/36 densovirus.•Purification yields infective, intact virion.•Pelleted virus can be re-suspended at desired concentration.

Dengue virus (DENV) is a mosquito-transmitted flavivirus that infects approximately 100 million people annually. Multi-day protocols for purification of DENV reduce the infective titer due to viral sensitivity to both temperature and pH. Herein we describe a 5-h protocol for the purification of all DENV serotypes, utilizing traditional gradient-free ultracentrifugation followed by selective virion precipitation. This protocol allows for the separation of DENV from contaminating proteins - including intact C6/36 densovirus, for the production of infective virus at high concentration for protein-level analysis.