[从噬菌体展示文库中筛选和鉴定抗日本血吸虫的噬菌体抗体]
[Screening and identification of phage antibodies from phage display library against Schistosoma japonicum].
作者信息
Chen Dai-xiong, Yu Mu-hua, Lei Zhi-gang, Zheng Bin, He Ai, Zhang Rui-lin, Zhan Xi-mei
机构信息
Department of Parasitology, Guangzhou Medical College, Guangzhou 510182.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2003;21(5):261-3.
OBJECTIVE
To obtain single chain variable fragment(ScFv) against the circulating antigen(CAg) from Schistosoma japonicum(Sj).
METHODS
Metabolic antigen of adult worm of Sj (Sj-MAg) was used in the panning of phage library against Schistosoma japonicum. The activity of Sj-MAg-binding phage clones was assayed by ELISA. The specificity of expression products of the positive clones was analyzed by ELISA, SDS-PAGE and Western blotting.
RESULTS
Seventy-two randomly selected clones were tested for the presence of anti-Sj-MAg ScFvs, 6 clones showed positive. The specificity of these 6 clones was confirmed by binding them to antigens of other four trematodes. Two clones (B04, C24) were found to bind to Sj-MAg but not to any of the antigens of other four trematodes and their expression products were about 31 kDa in size.
CONCLUSION
ScFv antibodies against the circulating antigens from Schistosoma japonicum Sj-MAg can be selected and manufactured from the antibody library.
目的
获取针对日本血吸虫循环抗原（CAg）的单链可变片段（ScFv）。
方法
用日本血吸虫成虫代谢抗原（Sj-MAg）对噬菌体文库进行筛选，以对抗日本血吸虫。通过酶联免疫吸附测定（ELISA）检测与Sj-MAg结合的噬菌体克隆的活性。用ELISA、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹法分析阳性克隆表达产物的特异性。
结果
对随机挑选的72个克隆进行抗Sj-MAg ScFv检测，6个克隆呈阳性。通过将这6个克隆与其他四种吸虫的抗原结合，证实了它们的特异性。发现两个克隆（B04、C24）能与Sj-MAg结合，但不与其他四种吸虫的任何抗原结合，其表达产物大小约为31 kDa。
结论
可从抗体文库中筛选并制备针对日本血吸虫Sj-MAg循环抗原的ScFv抗体。
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