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构建一个天然人单链可变片段(scFv)文库,用于鉴定抗伤寒沙门氏菌溶血素E抗原的单克隆scFv。

Generation of a naïve human single chain variable fragment (scFv) library for the identification of monoclonal scFv against Salmonella Typhi Hemolysin E antigen.

作者信息

Lim Bee Nar, Chin Chai Fung, Choong Yee Siew, Ismail Asma, Lim Theam Soon

机构信息

Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.

Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.

出版信息

Toxicon. 2016 Jul;117:94-101. doi: 10.1016/j.toxicon.2016.04.032. Epub 2016 Apr 14.

Abstract

Antibody phage display is a useful tool for the isolation and identification of monoclonal antibodies. Naive antibody libraries are able to overcome the limitations associated with the traditional hybridoma method for monoclonal antibody generation. Antibody phage display is also a preferred method for antibody generation against toxins as it does not suffer from toxicity mediated complications. Here, we describe a naïve multi ethnic scFv antibody library generated via two-step cloning with an estimated diversity of 2 × 10(9). The antibody library was used to screen for monoclonal antibodies against Hemolysin E antigen, a pore forming toxin produced by Salmonella enterica serovar Typhi. A soluble monoclonal scFv antibody against the HlyE toxin (IgM scFv D7 anti-hlyE) was isolated from the library. This shows the value of the naïve library to generate antibodies against toxin targets in addition to the potential use of the library to isolate antibodies against other immunogenic targets.

摘要

抗体噬菌体展示是用于分离和鉴定单克隆抗体的一种有用工具。天然抗体文库能够克服与传统杂交瘤方法生产单克隆抗体相关的局限性。抗体噬菌体展示也是针对毒素产生抗体的一种优选方法,因为它不会出现由毒性介导的并发症。在此,我们描述了一个通过两步克隆产生的具有估计多样性为2×10⁹的天然多民族单链抗体(scFv)文库。该抗体文库用于筛选针对溶血素E抗原的单克隆抗体,溶血素E抗原是由伤寒沙门氏菌血清型伤寒杆菌产生的一种成孔毒素。从该文库中分离出了一种针对HlyE毒素的可溶性单克隆scFv抗体(IgM scFv D7抗hlyE)。这表明了天然文库除了可用于分离针对其他免疫原性靶标的抗体外,对于产生针对毒素靶标的抗体的价值。

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