Amgen Research, Amgen Asia R&D Center, Amgen Biopharmaceutical R&D (Shanghai) Co., Ltd., 13th Floor, Building No. 2, 4560 Jinke Road, Zhangjiang, Shanghai, 201210, China.
Shanghai Public Health Clinical Center, Institutes of Biomedical Sciences, Fudan University, PI's Office IV, Scientific Research Center Building, 2901 Caolang Road, Jin Shan District, Shanghai, 201508, P. R. China.
Commun Biol. 2020 Mar 26;3(1):146. doi: 10.1038/s42003-020-0867-7.
Hybridoma and phage display are two powerful technologies for isolating target-specific monoclonal antibodies based on the binding. However, for complex membrane proteins, such as G protein-coupled receptors (GPCRs), binding-based screening rarely results in functional antibodies. Here we describe a function-based high-throughput screening method for quickly identifying antibody antagonists and agonists against GPCRs by combining glycosylphosphatidylinositol-anchored antibody cell display with β-arrestin recruitment-based cell sorting and screening. This method links antibody genotype with phenotype and is applicable to all GPCR targets. We validated this method by identifying a panel of antibody antagonists and an antibody agonist to the human apelin receptor from an immune antibody repertoire. In contrast, we obtained only neutral binders and antibody antagonists from the same repertoire by phage display, suggesting that the new approach described here is more efficient than traditional methods in isolating functional antibodies. This new method may create a new paradigm in antibody drug discovery.
杂交瘤和噬菌体展示是两种强大的技术，可基于结合来分离针对特定靶标的单克隆抗体。然而，对于复杂的膜蛋白，例如 G 蛋白偶联受体（GPCR），基于结合的筛选很少产生功能抗体。在这里，我们描述了一种基于功能的高通量筛选方法，该方法将糖基磷脂酰肌醇锚定的抗体细胞展示与β-arrestin 募集的细胞分选和筛选相结合，用于快速鉴定针对 GPCR 的抗体拮抗剂和激动剂。该方法将抗体基因型与表型联系起来，适用于所有 GPCR 靶标。我们通过从免疫抗体库中鉴定出一组针对人 Apelin 受体的抗体拮抗剂和抗体激动剂来验证该方法。相比之下，我们通过噬菌体展示仅从同一库中获得了中性结合物和抗体拮抗剂，这表明与传统方法相比，这里描述的新方法在分离功能抗体方面更加高效。这种新方法可能会在抗体药物发现中创造一个新的范例。
