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Lipid-tagged single domain antibodies for improved enzyme-linked immunosorbent assays.
Journal of Immunological Methods ( IF 2.2 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.jim.2020.112790
Lisa C Shriver-Lake 1 , Ellen R Goldman 1 , Scott N Dean 1 , Jinny L Liu 1 , Tamiya M Davis 2 , George P Anderson 1
Affiliation  

Anti-Staphylococcal Enterotoxin B single domain antibodies were engineered to include the N-terminal peptide sequence of the major outer membrane lipoprotein from Escherichia coli, which directs the N-terminal addition of lipid to the single domain antibody. We produced and purified two different single domain antibodies as well as a variant and dimer construct of one of the two, all with and without the added lipid. Their ability to function as the capture antibody in standard enzyme-linked immunosorbent assays were evaluated, finding that coating polystyrene microtiter plates with the lipid-tagged single domain antibodies gave a 3-fold improvement in the observed limit of detection. This increase was likely due to an increased amount of single domain antibody adsorbed to the microtiter plate, which translated to improved limits of detection of Staphylococcal Enterotoxin B over using the same single domain antibody sans lipid-tag. However, improved orientation may also play a role. Regardless of the mechanism, the biosynthetic lipid-tagging of single domain antibodies represent a facile modality that can enhance their ability to be utilized as immunoassay capture reagent as well as facilitate their incorporation into liposome targeting applications in the future.



中文翻译:

脂质标记的单域抗体,用于改进的酶联免疫吸附测定。

抗葡萄球菌肠毒素B单结构域抗体经过工程改造,以包含大肠杆菌主要外膜脂蛋白的N端肽序列,它指导脂质的N端添加到单结构域抗体中。我们生产并纯化了两种不同的单域抗体,以及两种抗体之一的变体和二聚体构建体,均带有或不带有添加的脂质。评估了它们在标准酶联免疫吸附测定法中充当捕获抗体的能力,发现用脂质标记的单结构域抗体包被聚苯乙烯微量滴定板可将观察到的检测限提高3倍。这种增加可能是由于吸附到微量滴定板上的单域抗体数量增加,与使用相同的单域抗体(不含脂质标签)相比,这意味着葡萄球菌肠毒素B的检测限得到了提高。但是,改善方向也可能起作用。不管机制如何

更新日期:2020-05-15
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