NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies
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DOI：
10.7554/elife.16228
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发表时间：
2016-07-19
期刊：
影响因子：
7.7
通讯作者：
Perez, Franck
中科院分区：
文献类型：
--
作者：
Moutel, Sandrine;Bery, Nicolas;Perez, Franck
抗体的体外选择允许快速且以较低成本获得高功能性结合剂。在这里，我们描述了第一个完全合成的人源化美洲驼单结构域抗体的噬菌体展示文库（NaLi-H1：Nanobody Library Humanized 1）。基于针对细胞内稳定性优化的人源化合成单结构域抗体（hs 2dAb）支架，高度多样化的文库提供高亲和力结合剂而无需动物免疫。NaLi-H1按照几种针对各种靶标（荧光蛋白、肌动蛋白、微管蛋白、p53、HP 1）的选择方案进行筛选。还获得了针对活性RHO GT3的构象抗体。选择的hs 2dAb用于各种免疫测定，并且通常被发现是功能性胞内抗体，能够追踪或抑制内源性靶标。胞内抗体的功能化允许活细胞中的特异性蛋白质敲低。最后，针对肿瘤细胞表面的直接选择产生了针对肿瘤特异性抗原的hs 2dAb，进一步突出了该文库用于治疗应用的潜在用途。
In vitro selection of antibodies allows to obtain highly functional binders, rapidly and at lower cost. Here, we describe the first fully synthetic phage display library of humanized llama single domain antibody (NaLi-H1: Nanobody Library Humanized 1). Based on a humanized synthetic single domain antibody (hs2dAb) scaffold optimized for intracellular stability, the highly diverse library provides high affinity binders without animal immunization. NaLi-H1 was screened following several selection schemes against various targets (Fluorescent proteins, actin, tubulin, p53, HP1). Conformation antibodies against active RHO GTPase were also obtained. Selected hs2dAb were used in various immunoassays and were often found to be functional intrabodies, enabling tracking or inhibition of endogenous targets. Functionalization of intrabodies allowed specific protein knockdown in living cells. Finally, direct selection against the surface of tumor cells produced hs2dAb directed against tumor-specific antigens further highlighting the potential use of this library for therapeutic applications.