Purification of adult Anguillicola crassus whole-worm extract antigens for detection of specific antibodies in serum from the European eel (Anguilla anguilla) by ELISA
Purification of adult Anguillicola crassus whole-worm extract antigens for detection of specific antibodies in serum from the European eel (Anguilla anguilla) by ELISA
复制标题
纯化成年鳗鲡全虫提取物抗原，用于通过 ELISA 检测欧洲鳗鱼 (Anguilla anguilla) 血清中的特异性抗体
DOI：
10.1006/fsim.1994.1027
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发表时间：
1994
影响因子：
4.7
通讯作者：
L. Pilström
中科院分区：
文献类型：
--
作者：
J. Höglund;L. Pilström
摘要以往的研究表明，厚壳鳗能激发鳗鱼的免疫反应。本研究采用间接酶联免疫吸附试验（ELISA）研究了鳗鲡对厚鳗鲡的体液免疫应答。成虫全虫提取物用作抗原，亲和纯化的多克隆兔抗鳗鱼IG抗体偶联碱性磷酸酶用作偶联物。测定了来自（1）未感染;（2）自然感染和（3）用全虫提取物免疫的鳗鱼的合并血清中的特异性抗体水平。使用该测定，注意到高背景水平，可能是由于成虫肠道中的鳗鱼免疫球蛋白污染了抗原。因此，在不同类型的合并血清之间观察到吸光度值的微小差异。然而，通过纯化抗原，包括亲和层析、凝胶过滤和等电聚焦技术，增加了测定的灵敏度，使得有可能区分不同类型血清之间的抗体水平。
Abstract Previous studies indicate that Anguillicola crassus elicits an immune response in the eel. In this study, the humoral immune response of the eel to Anguillicola crassus has been investigated using an indirect enzyme-linked immunosorbent assay (ELISA). Adult whole-worm extract was used as an antigen and affinity purified polyclonal rabbit anti-eel Ig antibodies coupled to alkaline phosphatase were used as a conjugate. The specific antibody levels in pooled sera from eels that were (1) non-infected; (2) naturally infected and (3) immunised with whole-worm extract, were measured. Using this assay, high background levels were noted, probably due to contamination of the antigen by eel immunoglobulin from the gut of the adult parasite. Consequently, small differences in absorbance values were noted between the different types of pooled sera. However, by purification of the antigen including affinity chromatography, gel filtration and isoelectric focusing techniques the sensitivity of the assay was increased, making it possible to differentiate the antibody levels between the different types of sera.