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铜绿假单胞菌外毒素:通过制备性聚丙烯酰胺凝胶电泳进行纯化及高效特异性抗毒素血清的研制

Pseudomonas aeruginosa exotoxin: purification by preparative polyacrylamide gel electrophoresis and the development of a highly specific antitoxin serum.

作者信息

Callahan L T

出版信息

Infect Immun. 1976 Jul;14(1):55-61. doi: 10.1128/iai.14.1.55-61.1976.

Abstract

Pseudomonas aeruginosa exotoxin has been purified to a specific activity of 12,000 to 16,000 mouse median lethal doses/mg of protein. Total recovery was about 25%, and the degree of purification was approximately 3,000-fold. Preparative polyacrylamide gel electrophoresis greatly facilitated purification. As judged by analytical disc gel electrophoresis, the purified toxin contained one major band of protein and only a negligible amount of contamination. Antiserum prepared against the purified toxin neutralized the lethal activity of crude toxin preprations and reacted by double immunodiffusion with a single component of concentrated broth cultures of P. aeruginosa isolates obtained from a clinical source.

摘要

铜绿假单胞菌外毒素已被纯化至比活性为12,000至16,000小鼠半数致死剂量/毫克蛋白质。总回收率约为25%,纯化程度约为3000倍。制备性聚丙烯酰胺凝胶电泳极大地促进了纯化过程。通过分析性圆盘凝胶电泳判断,纯化后的毒素含有一条主要的蛋白质条带,且污染量可忽略不计。用纯化毒素制备的抗血清可中和粗毒素制剂的致死活性,并通过双向免疫扩散与从临床来源获得的铜绿假单胞菌分离株浓缩肉汤培养物的单一成分发生反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8c53/420843/71066489340b/iai00223-0073-a.jpg

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