Ma J, Inzana T J
Department of Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg 24061.
Am J Vet Res. 1992 Jan;53(1):59-62.
An efficient, single-step method for purification of the 110-kilodalton (kDa) hemolysin of Actinobacillus pleuropneumoniae was developed. An immunoaffinity column was made by cross-linking murine monoclonal antibody 8C2 to the 110-kDa hemolysin of A pleuropneumoniae strain J45 serotype 5 to protein A-agarose beads. Purified hemolysin with high hemolytic activity was obtained after washing the column with phosphate-buffered saline solution, and eluting the hemolysin with 50 mM diethylamine, pH 11.0. The same column was also used to purify the hemolysin from A pleuropneumoniae strain 4074 serotype 1. The purification procedure could be completed within 5 hours, and almost 50% of the total hemolytic activity and hemolysin protein was recovered in pure form.
开发了一种高效的单步方法，用于纯化胸膜肺炎放线杆菌的110千道尔顿（kDa）溶血素。通过将鼠单克隆抗体8C2与胸膜肺炎放线杆菌J45血清型5的110-kDa溶血素交联到蛋白A-琼脂糖珠上，制成免疫亲和柱。用磷酸盐缓冲盐水溶液洗涤柱子，并用pH 11.0的50 mM二乙胺洗脱溶血素后，获得了具有高溶血活性的纯化溶血素。同一柱子也用于从血清型1的胸膜肺炎放线杆菌4074菌株中纯化溶血素。纯化过程可在5小时内完成，几乎50%的总溶血活性和溶血素蛋白以纯形式回收。
