[Isolation and determination of activity of IgA1 protease from Neisseria meningitidis].

Bioorganicheskaia khimiia Pub Date : 2010-01-01
E Iu Iagudaeva, L S Zhigis, O A Razguliaeva, V S Zueva, E E Mel'nikov, V P Zubov, L V Kozlov, A M Bichucher, O V Kotel'nikova, A P Alliluev, A E Avakov, L D Rumsh
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Abstract

A method of the isolation and purification of IgAl protease from a culture of Neisseria meningitidis serogroup A has been developed. Three inactivated intermediates of the production of the meningococcal vaccine, a culture liquid, as well as a supernatant and sediment obtained by the precipitation of bacterial cells by cetavlon, served as a starting material. The purity of IgA1 protease was determined by SDS-PAGE. An immunoenzyme assay for determining the IgA1 protease activity has been devised. The yield of the enzyme with a specific activity of 0.5 to 4 million units/mg from 103 g of the cetavlon precipitate (40 l of culture liquid) was about 600 mug. It was shown that IgAl protease isolated from serogroup A meningococcus is capable of protecting experimental animals (mice) infected with meningococcus of serogroup B.

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[脑膜炎奈瑟菌IgA1蛋白酶的分离及活性测定]。
建立了一种从脑膜炎奈瑟菌血清A群培养物中分离纯化IgAl蛋白酶的方法。三种生产脑膜炎球菌疫苗的灭活中间体,一种培养液,以及由cetavlon沉淀细菌细胞获得的上清液和沉淀物,作为起始材料。用SDS-PAGE法测定IgA1蛋白酶的纯度。设计了一种测定IgA1蛋白酶活性的免疫酶测定法。从103g的cetavlon沉淀物(40l培养液)中得到比活性为50 ~ 400万单位/mg的酶,产率约为600马克。结果表明,从A群脑膜炎球菌中分离的IgAl蛋白酶对感染B群脑膜炎球菌的实验动物(小鼠)具有保护作用。
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