Orr H C, Weetall H H, Probst P G, Littlejohn D C, Chu F C, Johnson J B, Petricciani J C
J Clin Microbiol. 1976 Apr;3(4):402-5. doi: 10.1128/jcm.3.4.402-405.1976.
Use of immunoadsorbents to remove bacteriophages from tissue culture serum was investigated. Immune globulins from rabbit antiserum prepared against phi V-1 phage were immobilized by covalent linkage to activated porous silica glass derivatives of p-aminoarylamine and to Sepharose-4B. Chromatographic columns of each material were used to filter samples of a fetal bovine serum into which had been introduced 8100 plaque-forming units of the phage per ml. Efficiency of removal was determined by plaque assays of phi V-1 phage recovered in the effluent fluids. Activated but uncoupled matrices nonspecifically removed from 49 to 59% of the phages introduced into the experimental serum. A reduction of 35 to 37% in phage content occurred in the serum after filtration through columns coupled to nonantibody protein. With specific immune globulins attached, the Sepharose-4B matrix reduced the concentration of phage in the serum below a detectable quantity. Noapparent alterations occurred in the growth-promoting property of serum filtered through the Sepharose-4B immunoadsorbent as measured by cloning efficiency of BHK-21, WI-38, and FRhL-2 cells. These experiments serve as a model system for use of immunoadsorbents for selective removal of bacteriophages and perhaps other extraneous microbial agents from tissue culture serum.
研究了使用免疫吸附剂从组织培养血清中去除噬菌体的方法。用针对φV - 1噬菌体制备的兔抗血清中的免疫球蛋白，通过共价连接固定到对氨基芳胺的活化多孔硅胶衍生物和琼脂糖-4B上。使用每种材料的色谱柱过滤胎牛血清样品，该血清中每毫升已引入8100个噬菌斑形成单位的噬菌体。通过对流出液中回收的φV - 1噬菌体进行噬菌斑测定来确定去除效率。活化但未偶联的基质非特异性地去除了引入实验血清中49%至59%的噬菌体。通过与非抗体蛋白偶联的柱过滤后，血清中的噬菌体含量降低了35%至37%。附着有特异性免疫球蛋白的琼脂糖-4B基质将血清中的噬菌体浓度降低到检测不到的量以下。通过BHK - 21、WI - 38和FRhL - 2细胞的克隆效率测量，通过琼脂糖-4B免疫吸附剂过滤的血清的生长促进特性没有明显改变。这些实验作为一个模型系统，用于使用免疫吸附剂从组织培养血清中选择性去除噬菌体以及可能的其他外来微生物制剂。
