Makino M, Ishikawa G, Yamaguchi K, Okada Y, Watanabe K, Sasaki-Iwaki Y, Manabe S, Honda M, Komuro K
Department of Bacterial and Blood Products, National Institute of Health, Tokyo, Japan.
Arch Virol. 1994;139(1-2):87-96. doi: 10.1007/BF01309456.
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.
体外实验需要浓缩的活逆转录病毒。一种最初为去除生物危害病毒而开发的铜铵再生纤维素中空纤维，称为BMM，被用于浓缩两种不同的逆转录病毒，即嗜亲性鼠白血病病毒（MuLV）和人类免疫缺陷病毒（HIV）。BMM可用于从500 - 1000毫升培养上清液中将活病毒悬液浓缩10至30倍。通过XC蚀斑形成试验和逆转录酶试验证明，经BMM浓缩的嗜亲性MuLV具有活性且生物学完整性良好。浓缩后的MuLV在小鼠脾脏中的滴度比原液高得多。通过p24抗原评估的HIV病毒浓度明显高于HIV感染细胞系的原始培养上清液。由于BMM中空纤维通过筛分机制捕获病毒而非吸附，通过洗涤可回收病毒颗粒，且活病毒总回收率较高，约为50%。此外，在过滤面积为0.03平方米的情况下，处理1000毫升上清液60分钟就足够了。这些结果表明，BMM为我们提供了一种快速、安全且高效的浓缩活逆转录病毒的方法。
