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用于鲤科鱼类卵黄蛋白原检测的新型重组单克隆抗体。

Novel recombinant monoclonal antibodies for vitellogenin assays in cyprinid fish species.

作者信息

Rao Yu, Zhong Liqiao, Liao Tao, Jin Shiwei, Wang Yuzhen, Song Bo, Li Jia, Zhang Xiaohua, Hemmingsen Sean M, Xu Ying, Dai Heping

机构信息

Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuchang, Wuhan, Hubei 430072, PR China.

出版信息

Dis Aquat Organ. 2010 Dec 7;93(1):83-91. doi: 10.3354/dao02268.

Abstract

Various polyclonal and monoclonal antibodies have been developed for vitellogenin (Vtg) bioassays in different aquatic species. Preparation of these reagents is time-consuming and expensive. In the present study, a phage-displayed, recombinant, single-chain variable fragment (scFv) format antibody library was constructed using splenic mRNA from non-immunized mice. After 3 rounds of panning, 3 scFv antibodies with specificity for the highly conserved N-terminal region of cyprinid fish Vtg were isolated. One of these, antibody H4, bound purified Vtg from common carp Cyprinus carpio, zebrafish Danio rerio and Chinese rare minnow Gobiocypris rarus with similar affinities and detected Vtg in zebrafish plasma samples. This study provides a simple, low cost Vtg bioassay for plasma samples from a variety of cyprinid fish.

摘要

已开发出多种用于不同水生生物卵黄蛋白原(Vtg)生物测定的多克隆和单克隆抗体。这些试剂的制备既耗时又昂贵。在本研究中,使用未免疫小鼠的脾脏mRNA构建了噬菌体展示的重组单链可变片段(scFv)形式的抗体文库。经过三轮淘选,分离出了3种对鲤科鱼类Vtg高度保守的N端区域具有特异性的scFv抗体。其中一种抗体H4,以相似的亲和力结合鲤鱼、斑马鱼和稀有鮈鲫的纯化Vtg,并能检测斑马鱼血浆样本中的Vtg。本研究为多种鲤科鱼类的血浆样本提供了一种简单、低成本的Vtg生物测定方法。

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