Liu Jinny L, Shriver-Lake Lisa C, Zabetakis Dan, Goldman Ellen R, Anderson George P
Naval Research Laboratory, Center for Biomolecular Science and Engineering, Washington, DC 20375, USA.
Antibodies (Basel). 2018 Dec 15;7(4):44. doi: 10.3390/antib7040044.
In this work, we describe the selection and characterization of single-domain antibodies (sdAb) towards the E2/E3E2 envelope protein of the Western equine encephalitis virus (WEEV). Our purpose was to identify novel recognition elements which could be used for the detection, diagnosis, and perhaps treatment of western equine encephalitis (WEE). To achieve this goal, we prepared an immune phage display library derived from the peripheral blood lymphocytes of a llama that had been immunized with an equine vaccine that includes killed WEEV (West Nile Innovator + VEWT). This library was panned against recombinant envelope (E2/E3E2) protein from WEEV, and seven representative sdAb from the five identified sequence families were characterized. The specificity, affinity, and melting point of each sdAb was determined, and their ability to detect the recombinant protein in a MagPlex sandwich immunoassay was confirmed. Thus, these new binders represent novel recognition elements for the E2/E3E2 proteins of WEEV that are available to the research community for further investigation into their applicability for use in the diagnosis or treatment of WEE.
在这项工作中，我们描述了针对西部马脑炎病毒（WEEV）E2/E3E2包膜蛋白的单域抗体（sdAb）的筛选和表征。我们的目的是鉴定可用于检测、诊断以及可能用于治疗西部马脑炎（WEE）的新型识别元件。为实现这一目标，我们制备了一个免疫噬菌体展示文库，该文库源自一只经包含灭活WEEV（西尼罗河创新株+VEWT）的马用疫苗免疫的美洲驼外周血淋巴细胞。该文库用来自WEEV的重组包膜（E2/E3E2）蛋白进行淘选，并对来自五个已鉴定序列家族的七个代表性sdAb进行了表征。测定了每个sdAb的特异性、亲和力和熔点，并证实了它们在磁珠夹心免疫测定中检测重组蛋白的能力。因此，这些新的结合物代表了WEEV的E2/E3E2蛋白的新型识别元件，可供研究界进一步研究它们在WEE诊断或治疗中的适用性。
