A gradient-free method for the purification of infective dengue virus for protein-level investigations.
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.
The Department of Chemistry and Biochemistry, The University of Arizona, 1306 E. University Blvd., Tucson, AZ 85721, USA.
Stephanie M
SM
Jensen
The Department of Chemistry and Biochemistry, The University of Arizona, 1306 E. University Blvd., Tucson, AZ 85721, USA.
Celina T
CT
Nguyen
The Department of Chemistry and Biochemistry, The University of Arizona, 1306 E. University Blvd., Tucson, AZ 85721, USA. Electronic address: jjewett@email.arizona.edu.
John C
JC
Jewett
D000330:Aedes / Q000821:virology; D000818:Animals; D002460:Cell Line; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000145:classification / Q000302:isolation & purification* / Q000502:physiology; D017990:Densovirus / Q000302:isolation & purification; D000072138:Mosquito Vectors / Q000821:virology; D065288:Serogroup; D013997:Time Factors; D014461:Ultracentrifugation; D014773:Virology / Q000379:methods*; D014779:Virus Replication
Aedes albopictus; Dengue virus; Densovirus; Infectivity; Precipitation; Purification