Alphavirus Purification Using Low-speed Spin Centrifugation

Vamseedhar Rayaprolu; Jolene Ramsey; Joseph Che-Yen Wang; Suchetana Mukhopadhyay

doi:10.21769/BioProtoc.2772

Alphavirus Purification Using Low-speed Spin Centrifugation

Bio Protoc. 2018 Mar 20;8(6):e2772. doi: 10.21769/BioProtoc.2772.

Authors

Vamseedhar Rayaprolu¹, Jolene Ramsey¹, Joseph Che-Yen Wang¹, Suchetana Mukhopadhyay¹

Affiliation

¹ Department of Biology, Indiana University, Bloomington, IN, USA.

Abstract

Chemical and sedimentation procedures are used to purify virus particles. While these approaches are successful for wild-type viruses, they are often not feasible for purifying mutant viruses with assembly defects. We combined two published methods ( Atasheva et al., 2013 ; Moller- Tank et al., 2013 ), to generate a protocol that uses low-speed centrifugation to purify both wildtype and mutant enveloped virus particles at high yield with minimal handling steps. This protocol has successfully been used to purify alphavirus particles for imaging and structural studies ( Wang et al., 2015 ; Ramsey et al., 2017 ).

Keywords: Assembly mutants; Centrifugation; Enveloped virus purification.