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通过过滤色谱法浓缩噬菌体裂解物。

Concentration of bacteriophage lysates by filter chromatography.

作者信息

Goyal S M, Zerda K S, Gerba C P

出版信息

J Virol Methods. 1980;1(2):79-85. doi: 10.1016/0166-0934(80)90016-6.

Abstract

High-titered phage stocks are required for carrying out genetic and physicochemical studies on bacteriophages. This study describes a simple method for the concentration of coliphages MS-2 and T2 by charge-modified filters. Phage lysates were first clarified by filtration through serum-coated membrane filters. The clarified lysate was adjusted to pH 6 and passed through a Zeta-plus filter (30 S size). Greater than 99% virus adsorption occurred under these conditions. Adsorbed viruses were successfully eluted by small volumes of 3% beef extract (pH 10) which was subsequently neutralized with 3% beef extract (pH2). A concentration factor as great as 126 could be achieved by this method, with recoveries ranging from 54% to 97% (average = 60%).

摘要

进行噬菌体的遗传和物理化学研究需要高滴度的噬菌体储备液。本研究描述了一种通过电荷修饰滤器浓缩大肠杆菌噬菌体MS - 2和T2的简单方法。噬菌体裂解液首先通过血清包被的膜滤器过滤以澄清。将澄清的裂解液调节至pH 6,然后通过Zeta-plus滤器(30 S尺寸)。在这些条件下,病毒吸附率大于99%。吸附的病毒用少量3%牛肉浸出液(pH 10)成功洗脱,随后用3%牛肉浸出液(pH 2)中和。通过该方法可实现高达126的浓缩系数,回收率在54%至97%之间(平均 = 60%)。

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