Lauer Björn, Ottleben Ilka, Jacobsen Hans-Jörg, Reinard Thomas
LG Molekulargenetik, University of Hannover, Herrenhäuser Strasse 2, D-30419 Hannover, Germany.
J Agric Food Chem. 2005 Feb 23;53(4):899-904. doi: 10.1021/jf048651s.
The selection of synthetic antibody fragments from large phage libraries has become a common method for the generation of specific antibodies. The technique is particularly valuable when antibodies against 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 is not detectable. Here, a single variable chain antibody (scFv) against the highly toxic mycotoxin fumonisin B1 has been produced. The hapten was coupled via a linker to biotin. Using this conjugate and a naive scFv library, it was possible to circumvent both the necessity of immunization and the risk of a disguised hapten. The scFv obtained after three panning rounds was found to bind specifically to 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 antibody obtained can potentially be used for developing a rapid and affordable immunoassay for detection of food contamination and can be applied in immunoaffinity chromatography, usually carried out prior to HPLC analysis of mycotoxin-contaminated food and feed.
从大型噬菌体文库中筛选合成抗体片段已成为产生特异性抗体的常用方法。当必须生产针对小的、无免疫原性的分子（半抗原）或剧毒物质的抗体时，该技术特别有价值。此外，半抗原通常与蛋白质载体偶联，存在游离半抗原无法检测到的风险。在此，已产生了一种针对剧毒霉菌毒素伏马菌素B1的单链可变区抗体（scFv）。半抗原通过接头与生物素偶联。使用这种缀合物和一个未经免疫的scFv文库，既可以避免免疫的必要性，又可以避免半抗原被掩盖的风险。经过三轮淘选后获得的scFv被发现能特异性结合游离的伏马菌素B1和伏马菌素-生物素缀合物。伏马菌素B2也能被该scFv结合。对scFv和伏马菌素B1分子的建模显示结构匹配良好。所获得的抗体有可能用于开发一种快速且经济的免疫测定方法以检测食品污染，并且可应用于免疫亲和色谱法，该方法通常在对受霉菌毒素污染的食品和饲料进行HPLC分析之前进行。
