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首页> 外文会议>Smart biomedical and physiological sensor technologies VII >Using llama derived single domain antibodies to target botulinum neurotoxins
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Using llama derived single domain antibodies to target botulinum neurotoxins

机译：使用美洲驼衍生的单结构域抗体靶向肉毒杆菌神经毒素

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Llama serum contains both conventional IgG as well as unique forms of antibody that contain only heavy chains where antigen binding is mediated through a single variable domain. These variable domains can be expressed recombinantly and are referred to as single domain antibodies (sdAb). SdAb are among the smallest known naturally derived antigen binding fragments, possess good solubility, thermal stability, and can refold after heat and chemical denaturation. Llamas were immunized with either BoNT A or B toxoid and phage display libraries prepared. Single domain antibodies (sdAb) that were able to detect botulinum neurotoxin (BoNT) serotypes A and B were selected from their respective libraries. Here, the binders obtained by panning the BoNT B library on either BoNT B toxoid or BoNT B complex toxoid coated plates or BoNT B toxin coupled microspheres are described. Using these panning methods, we selected for binders that showed specificity for BoNT B. Phage displayed binders were screened, moved to a protein expression vector and soluble sdAb was produced. Using a Luminex flow cytometer binders were evaluated in direct binding assays. We have exploited the unique properties of sdAb and used them as biological recognition elements in immuno-based sensors that can detect BoNT B.
机译：骆马血清既包含常规IgG，又包含独特形式的抗体，该抗体仅包含重链，其中抗原结合是通过单个可变域介导的。这些可变域可以重组表达，并称为单域抗体（sdAb）。 SdAb是已知的最小的天然来源抗原结合片段，具有良好的溶解性，热稳定性，并且在加热和化学变性后可以重新折叠。用BoNT A或B类毒素免疫骆驼，并制备噬菌体展示文库。从它们各自的文库中选择能够检测肉毒杆菌神经毒素（BoNT）血清型A和B的单域抗体（sdAb）。在此，描述了通过在BoNT B类毒素或BoNT B复合类毒素涂层平板或BoNT B毒素偶联的微球上淘选BoNT B文库而获得的粘合剂。使用这些淘选方法，我们选择了对BoNT B具有特异性的结合物。筛选出噬菌体展示的结合物，将其转移到蛋白质表达载体中，并产生可溶性sdAb。使用Luminex流式细胞仪，在直接结合测定中评估结合剂。我们已利用sdAb的独特特性，并将其用作可检测BoNT B的基于免疫的传感器中的生物识别元件。

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