Discovery of antibodies targeting multipass transmembrane proteins using a suspension cell-based evolutionary approach.
Discovery of antibodies targeting multipass transmembrane proteins using a suspension cell-based evolutionary approach.
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使用基于悬浮细胞的进化方法发现针对多次跨膜蛋白的抗体。
DOI：
10.1016/j.crmeth.2023.100429
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Spangler,JamieB
中科院分区：
文献类型：
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作者：
Krohl,PatrickJ;Fine,Justyn;Yang,Huilin;VanDyke,Derek;Ang,Zhiwei;Kim,KookBum;Thomas-Tikhonenko,Andrei;Spangler,JamieB
由于它们在细胞传感和信号处理中的关键功能，膜蛋白作为药理学靶标是高度优选的，并且抗体药物构成了医药市场上增长最快的治疗剂类别。然而，在开发识别复杂的多通道跨膜蛋白（如G蛋白偶联受体（GPCR））的抗体方面存在主要限制。这些挑战，主要是由于重组表达的多通道跨膜蛋白的困难，可以克服使用全细胞筛选技术，这使得功能性抗原在其天然构象的介绍。在这里，我们开发了基于悬浮细胞的全细胞淘选方法，以筛选针对天然酵母展示抗体库内的GPCR的特异性结合剂。我们实施了发现针对四种不同GPCR靶蛋白的高亲和力抗体的策略，证明了我们基于细胞的筛选工作流程推进靶向膜蛋白的抗体治疗剂发现的潜力。
Due to their critical functions in cell sensing and signal processing, membrane proteins are highly preferred as pharmacological targets, and antibody drugs constitute the fastest growing category of therapeutic agents on the pharmaceutical market. However, major limitations exist in developing antibodies that recognize complex, multipass transmembrane proteins, such as G-protein-coupled receptors (GPCRs). These challenges, largely due to difficulties with recombinant expression of multipass transmembrane proteins, can be overcome using whole-cell screening techniques, which enable presentation of the functional antigen in its native conformation. Here, we developed suspension cell-based whole-cell panning methodologies to screen for specific binders against GPCRs within a naive yeast-displayed antibody library. We implemented our strategy to discover high-affinity antibodies against four distinct GPCR target proteins, demonstrating the potential for our cell-based screening workflow to advance the discovery of antibody therapeutics targeting membrane proteins.