Generation of a highly diverse panel of antagonistic chicken monoclonal antibodies against the GIP receptor
Generation of a highly diverse panel of antagonistic chicken monoclonal antibodies against the GIP receptor
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DOI：
10.1080/19420862.2016.1276683
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发表时间：
2017-01-01
期刊：
影响因子：
5.3
通讯作者：
Izquierdo, Shelley
中科院分区：
文献类型：
--
作者：
Koenitzer, Jennifer D.;Pramanick, Shreya;Izquierdo, Shelley
产生针对G蛋白偶联受体（GPCR）的功能性抗体是具有挑战性的，这是由于它们的低密度表达、在不存在细胞膜的脂质双层的情况下的不稳定性以及可以充当抗原的通常短的胞外结构域。此外，特定的治疗概念可能需要抗体不仅结合受体，而且还充当功能性受体激动剂或拮抗剂。拮抗葡萄糖依赖性促胰岛素多肽（GIP）受体可能会在糖尿病和肥胖症的治疗中开辟新的治疗模式。因此，一组单克隆拮抗性抗体将是体外和体内概念验证研究的有用工具。该受体在啮齿动物和人类之间高度保守，这导致之前的小鼠和大鼠免疫活动产生很少的可用抗体。将免疫宿主切换到鸡，这是从哺乳动物遗传学上遥远的，使得能够产生含有172个独特序列的大而多样的单克隆抗体组。四分之三的鸡源性抗体的功能拮抗剂，表现出高亲和力的受体胞外结构域和采样广泛的表位库。对于困难的靶标，包括GPCR如GIPR，鸡正在成为用于治疗性抗体发现的有价值的免疫宿主。
Raising functional antibodies against G protein-coupled receptors (GPCRs) is challenging due to their low density expression, instability in the absence of the cell membrane's lipid bilayer and frequently short extracellular domains that can serve as antigens. In addition, a particular therapeutic concept may require an antibody to not just bind the receptor, but also act as a functional receptor agonist or antagonist. Antagonizing the glucose-dependent insulinotropic polypeptide (GIP) receptor may open up new therapeutic modalities in the treatment of diabetes and obesity. As such, a panel of monoclonal antagonistic antibodies would be a useful tool for in vitro and in vivo proof of concept studies. The receptor is highly conserved between rodents and humans, which has contributed to previous mouse and rat immunization campaigns generating very few usable antibodies. Switching the immunization host to chicken, which is phylogenetically distant from mammals, enabled the generation of a large and diverse panel of monoclonal antibodies containing 172 unique sequences. Three-quarters of all chicken-derived antibodies were functional antagonists, exhibited high-affinities to the receptor extracellular domain and sampled a broad epitope repertoire. For difficult targets, including GPCRs such as GIPR, chickens are emerging as valuable immunization hosts for therapeutic antibody discovery.