Production of a Single-Chain Variable Fragment Antibody against Fumonisin B1
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- 作者：Bjö ; rn Lauer ; Ilka Ottleben ; Hans-Jö ; rg Jacobsen ; and Thomas Reinard
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2005
- 出版时间：February 23, 2005
- 年：2005
- 卷：53
- 期：4
- 页码：899 - 904
- 全文大小：137K
- 年卷期：v.53,no.4(February 23, 2005)
- ISSN：1520-5118
文摘
The selection of synthetic antibody fragments from large phage libraries has become a commonmethod for the generation of specific antibodies. The technique is particularly valuable when antibodiesagainst small, non-immunogenic molecules (haptens) or highly toxic substances have to be produced.In addition, haptens are usually coupled to protein carriers, bearing the risk that the free hapten isnot detectable. Here, a single variable chain antibody (scFv) against the highly toxic mycotoxinfumonisin B1 has been produced. The hapten was coupled via a linker to biotin. Using this conjugateand a naïve scFv library, it was possible to circumvent both the necessity of immunization and therisk of a disguised hapten. The scFv obtained after three panning rounds was found to bind specificallyto both free fumonisin B1 and fumonisin-biotin conjugate. Also fumonisin B2 was bound by the scFv.Modeling of both scFv and fumonisin B1 molecule revealed a good fitting of structures. The antibodyobtained can potentially be used for developing a rapid and affordable immunoassay for detection offood contamination and can be applied in immunoaffinity chromatography, usually carried out priorto HPLC analysis of mycotoxin-contaminated food and feed.Keywords: ScFv; fumonisin; recombinant antibody; hapten; mycotoxin