Traul K, Larson D, Stephens R, Wolff J, Munch K, Mayyasi S
J Clin Microbiol. 1975 Sep;2(3):253-60. doi: 10.1128/jcm.2.3.253-260.1975.
Conventional methods (i.e. gradient centrifugation) for the purification of oncornaviruses are usually not effective in complete removal of nonviral proteins. Such contaminants often prove to be a nuisance in subsequent immunological or biochemical studies. Hyperimmune sera prepared from these viruses must be absorbed to assure specificity; cell-derived proteins can be shown to interfere with studies of virus structural proteins, nucleic acids, or viral enzymes. Herein is described a method for removal of most of these contaminants. Viruses are diluted in a high concentration of NaCl to achieve a final concentration of 15%, incubated for 30 min, sedimented, and resuspended in buffer. This procedure results in reductions of up to 48% of the protein without affecting particle count. Immunological, biochemical, and biological properties are not adversely affected. Of the proteins removed, fetal calf serum components and a ribonuclease (presumably cell-derived) were identified. This technique differs significantly from other high-salt methods in that the virus is not precipitated from suspension. It is believed that absorbed proteins are desorbed and left in solution (or suspension) as the virus is sedimented by centrifugation.
用于纯化肿瘤病毒的传统方法（即梯度离心法）通常无法有效完全去除非病毒蛋白。在后续的免疫学或生化研究中，这类污染物常常会造成麻烦。用这些病毒制备的超免疫血清必须经过吸收处理以确保特异性；已证实细胞衍生蛋白会干扰病毒结构蛋白、核酸或病毒酶的研究。本文描述了一种去除大部分这类污染物的方法。将病毒在高浓度氯化钠中稀释，使其最终浓度达到15%，孵育30分钟，沉淀后再重悬于缓冲液中。此步骤可使蛋白减少多达48%，而不影响病毒颗粒数量。免疫学、生化和生物学特性均未受到不利影响。在被去除的蛋白中，鉴定出了胎牛血清成分和一种核糖核酸酶（可能源自细胞）。该技术与其他高盐方法有显著不同，因为病毒不会从悬浮液中沉淀出来。据信，随着病毒通过离心沉淀，被吸附的蛋白会解吸并留在溶液（或悬浮液）中。
