Kopertekh Lilya, Meyer Torsten, Freyer Cornelia, Hust Michael
Julius Kühn-Institut, Bundesforschungsinstitut für Kulturpflanzen, Institut für die Sicherheit biotechnologischer Verfahren bei Pflanzen, Erwin-Baur-Str. 27, 06484, Quedlinburg, Germany.
Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr. 7, 38106, Braunschweig, Germany.
Biotechnol Rep (Amst). 2019 Feb 12;21:e00314. doi: 10.1016/j.btre.2019.e00314. eCollection 2019 Mar.
Typhimurium is one of the most important zoonotic pathogens worldwide and a major cause of economic losses in the pig production chain. The emergence of multi-drug resistant strains over the past years has led to considerations about an enhanced surveillance of bacterial food contamination. Currently, ELISA is the method of choice for high throughput identification of Typhimurium. The sensitivity and specificity of this assay might be improved by application of new diagnostic antibodies. We focused on plant-based expression of candidate diagnostic TM43-E10 antibodies discovered using as antigen the Typhimurium OmpD protein. The scFv-TM43-E10 and scFv-Fc-TM43-E10 antibody derivatives have been successfully produced in N. benthamiana using a deconstructed movement-deficient PVX vector supplemented with the γb silencing suppressor from Poa semilatent virus. The plant-made antibodies showed the same antigen-binding specificity as that of the microbial/mammalian cell-produced counterparts and could recognize the OmpD antigen in Typhimurium infected plant samples.
鼠伤寒沙门氏菌是全球最重要的人畜共患病原体之一，也是猪生产链中经济损失的主要原因。过去几年中多重耐药菌株的出现促使人们考虑加强对细菌食品污染的监测。目前，酶联免疫吸附测定（ELISA）是高通量鉴定鼠伤寒沙门氏菌的首选方法。应用新的诊断抗体可能会提高该检测方法的灵敏度和特异性。我们专注于基于植物表达候选诊断性TM43-E10抗体，该抗体以鼠伤寒沙门氏菌外膜蛋白D（OmpD）作为抗原被发现。单链抗体片段（scFv）-TM43-E10和单链抗体片段-免疫球蛋白Fc段（scFv-Fc）-TM43-E10抗体衍生物已通过使用补充了来自早熟禾半潜病毒γb沉默抑制子的解构运动缺陷型马铃薯X病毒（PVX）载体在本氏烟草中成功生产。植物源抗体表现出与微生物/哺乳动物细胞产生的对应抗体相同的抗原结合特异性，并且能够识别感染鼠伤寒沙门氏菌的植物样品中的OmpD抗原。
