Concentration of live retrovirus with a regenerated cellulose hollow fiber, BMM.
A concentrated live retrovirus is required for in vitro experiments. A cuprammonium-regenerated cellulose hollow fiber, termed BMM, originally developed for biohazardous viral removal, was used to concentrate two different retroviruses, an ecotropic murine leukemia virus (MuLV) and human immunodeficiency virus (HIV). The BMM was useful for concentrating live virus suspension 10- to 30-fold from 500-1000 ml of culture supernatant. The ecotropic MuLV concentrated by BMM was demonstrated to be viable and biologically intact by XC plaque-forming assay and reverse transcriptase assay. The concentrated MuLV reached a much higher titer in the spleen in mice than the original one. The virus concentration assessed by p24 antigen for HIV was clearly higher than that of the original culture supernatant of HIV-infected cell lines. Since BMM hollow fibers trapped viruses by the sieving mechanism but not by adsorption, the viral particles were recovered by washing and the total live virus recovery rate was high, about 50%. Furthermore 60 min sufficed to handle 1000 ml of supernatant in the case of a filtration area of 0.03 m2. These results show that the BMM provides us with a rapid, safe and efficient method for concentrating live retroviruses.
Makino
M
M
Department of Bacterial and Blood Products, National Institute of Health, Tokyo, Japan.
Ishikawa
G
G
Yamaguchi
K
K
Okada
Y
Y
Watanabe
K
K
Sasaki-Iwaki
Y
Y
Manabe
S
S
Honda
M
M
Komuro
K
K
D000818:Animals; D002460:Cell Line; D002482:Cellulose / Q000031:analogs & derivatives; D006678:HIV / Q000302:isolation & purification* / Q000502:physiology; D016655:HIV Core Protein p24 / Q000032:analysis; D007700:Kinetics; D009052:Leukemia Virus, Murine / Q000302:isolation & purification* / Q000502:physiology; D051379:Mice; D008854:Microscopy, Electron; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D013154:Spleen / Q000821:virology; D010948:Viral Plaque Assay; D014779:Virus Replication
D016655:HIV Core Protein p24; D002482:Cellulose; C009865:cuprammonium cellulose; D012194:RNA-Directed DNA Polymerase