...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Isolation of species-specific protein antigens of Rickettsia typhi and Rickettsia prowazekii for immunodiagnosis and immunoprophylaxis.
【24h】

Isolation of species-specific protein antigens of Rickettsia typhi and Rickettsia prowazekii for immunodiagnosis and immunoprophylaxis.

机译：伤寒立克次体和原立氏立克次体物种特异性蛋白抗原的分离，用于免疫诊断和免疫预防。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A simple procedure for the selective isolation of the protective species-specific protein antigens (SPAs) of Rickettsia typhi and Rickettsia prowazekii was developed to permit use of the SPAs in the immunodiagnosis and immunoprophylaxis of typhus infections. Although the SPAs were readily extracted from lysozyme- or detergent-treated rickettsiae, as measured by rocket immunoelectrophoresis, other polypeptides were also present, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In contrast, both water and seven buffers, each at a 10 mM concentration and pH 7.6, were nearly equally effective in the selective release of the SPAs from whole cells by extraction for 30 min at 45 degrees C. High-ionic-strength buffers and MgCl2 abolished this SPA release, thus suggesting that divalent cations were important in the binding of the SPAs to the cell envelope. The efficacy of the dilute buffer extraction procedure for isolation of large amounts of SPAs was tested by further characterization of the supernatants obtained by centrifugation (200,000 x g) of two successive tris-(hydroxymethyl)aminomethane-hydrochloride buffer (Tris) extracts. With this procedure, between 10 and 15 mg of SPA was obtained from 100 mg of purified rickettsiae. Although low-molecular-weight ribonucleic acid fragments were released into the Tris extracts in significant amounts, only the SPAs were detected, in significant quantities, as measured by polyacrylamide gel electrophoresis and rocket immunoelectrophoresis. The Tris extracts contained the same major and minor SPA polypeptides as those observed previously in SPA preparations obtained by extensive diethylaminoethyl-cellulose column chromatography, but the Tris SPAs were more satisfactory antigens in an enzyme-linked immunosorbent assay.
机译：开发了一种选择性分离伤寒立克次体和普氏立克次体的保护性物种特异性蛋白抗原（SPA）的简单程序，以允许将SPA用于斑疹伤寒感染的免疫诊断和免疫预防。尽管通过火箭免疫电泳测量很容易从溶菌酶或去污剂处理过的立克次体中提取出SPA，但也存在其他多肽，如十二烷基硫酸钠-聚丙烯酰胺凝胶电泳所示。相比之下，水和7种缓冲液（浓度分别为10 mM和pH 7.6）在45°C下提取30分钟从全细胞中选择性释放SPA的效果几乎相同。高离子强度缓冲液和MgCl2取消了该SPA释放，因此表明二价阳离子在SPA与细胞包膜的结合中很重要。通过进一步表征两个连续的三-（羟甲基）氨基甲烷-盐酸缓冲液（Tris）提取物的离心分离（200,000 x g）获得的上清液的特性，测试了稀释缓冲液提取程序分离大量SPA的功效。通过该程序，从100 mg纯化的立克次体中获得10至15 mg的SPA。尽管低分子量核糖核酸片段大量释放到Tris提取物中，但通过聚丙烯酰胺凝胶电泳和火箭免疫电泳检测到，只有大量SPA被检测到。 Tris提取物所含的主要和次要SPA多肽与以前通过大量二乙氨基乙基纤维素柱色谱法获得的SPA制剂中观察到的相同，但在酶联免疫吸附测定中，Tris SPA是更令人满意的抗原。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号