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用于免疫诊断的多房棘球绦虫各种重组抗原的特性分析。

Characterization of various recombinant antigens from Echinococcus multilocularis for use in the immunodiagnosis.

作者信息

Kouguchi Hirokazu, Suzuki Tomohiro, Yamano Kimiaki, Honma Hiroshi, Sawada Yukiharu

机构信息

Hokkaido Institute of Public Health, Kita-Ku, Sapporo 060-0819, Japan.

出版信息

Protein J. 2005 Jan;24(1):57-64. doi: 10.1007/s10930-004-0606-9.

Abstract

Using four clones isolated from Echinococcus multilocularis cDNA library with alveolar echinococcosis (AE) patient sera, various antigens were expressed as ThioHis tag-fused protein. Recombinant EmII/3 antigen was produced as the five fragments divided into the N-terminal (#5 and #5s), the central (#6 and #6s) and the C-terminal domain (#7). Immunoblot analysis revealed that the #7 showed significant reactivity whereas those of #5 and #5s were relatively low. The #6 and #6s also showed lower reactivity than that of #7, although the two minor bands of #6 reacted with every serum. These results suggested that an immunodominant region of EmII/3 locate within the C-terminal one third. The #8s recombinant antigen, Ser23-Glu176 of actin filament fragmenting protein (AFFP), apparently reacted with the AE patient sera, while the #1 antigen synthesized as a full-length antigen BI did not show such high reactivity. Thus, #7 and #8s antigens showed significant potential for use in immunodetection of AE. In addition, the specific antibodies against #7 and #8s reacted with specific antigens in crude extract of E. multilocularis cyst, indicating that these antigens retained antigenicity common to native EmII/3 and AFFP, respectively.

摘要

利用从多房棘球绦虫cDNA文库中分离出的四个克隆与泡型包虫病(AE)患者血清,将各种抗原表达为硫氧还蛋白组氨酸标签融合蛋白。重组EmII/3抗原作为五个片段产生,分为N端(#5和#5s)、中央(#6和#6s)和C端结构域(#7)。免疫印迹分析显示,#7表现出显著的反应性,而#5和#5s的反应性相对较低。#6和#6s的反应性也低于#7,尽管#6的两条小带与每份血清都有反应。这些结果表明,EmII/3的免疫显性区域位于C端的三分之一处。#8s重组抗原,肌动蛋白丝断裂蛋白(AFFP)的Ser23-Glu176,明显与AE患者血清发生反应,而作为全长抗原BI合成的#1抗原未表现出如此高的反应性。因此,#7和#8s抗原在AE免疫检测中显示出显著的应用潜力。此外,针对#7和#8s的特异性抗体与多房棘球绦虫囊肿粗提物中的特异性抗原发生反应,表明这些抗原分别保留了天然EmII/3和AFFP共有的抗原性。

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