Development of an Antigen-Antibody Co-Display System for Detecting Interaction of G-Protein-Coupled Receptors and Single-Chain Variable Fragments.
Development of an Antigen-Antibody Co-Display System for Detecting Interaction of G-Protein-Coupled Receptors and Single-Chain Variable Fragments.
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开发用于检测 G 蛋白偶联受体和单链可变片段相互作用的抗原抗体共展示系统
DOI：
10.3390/ijms22094711
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发表时间：
2021-04-29
影响因子：
5.6
通讯作者：
Li J
中科院分区：
文献类型：
--
作者：
Zhang Y;Wu BJ;Yu X;Luo P;Ye H;Yu Y;Han W;Li J
G蛋白偶联受体（GPCR），特别是趋化因子受体，是单克隆抗体药物的理想靶点。考虑到GPCR特殊的多通道跨膜结构，获得关于脱靶和抗原表位的抗体信息通常是一项繁重的工作。为了加快这一进程，需要开发一种快速简单的方法。基于分裂泛素的酵母双杂交系统（YTH）被用作新方法的蓝脚本。通过与跨膜肽融合，scFv抗体被设计成锚定在细胞膜上，其中GPCR也被共展示。偶联的split-ubiquitin系统将scFv-GPCR相互作用信号转化为报告基因的表达。通过优化单链抗体融合蛋白的拓扑结构和关键元件（包括信号肽、跨膜肽和柔性接头），建立了抗原-抗体共展示系统（Antigen-Antibody Co-Display，AACD），可快速检测抗体与靶GPCR（CXCR 4和CXCR 5）之间的相互作用，同时确定脱靶抗体和抗体相关表位。AACD系统可以快速确定GPCR与其候选抗体之间的关联，缩短脱靶检测和表位鉴定的研究周期。该系统将改进GPCR抗体的开发过程，为GPCR抗体的筛选提供新的策略。
G-protein-coupled receptors (GPCRs), especially chemokine receptors, are ideal targets for monoclonal antibody drugs. Considering the special multi-pass transmembrane structure of GPCR, it is often a laborious job to obtain antibody information about off-targets and epitopes on antigens. To accelerate the process, a rapid and simple method needs to be developed. The split-ubiquitin-based yeast two hybrid system (YTH) was used as a blue script for a new method. By fusing with transmembrane peptides, scFv antibodies were designed to be anchored on the cytomembrane, where the GPCR was co-displayed as well. The coupled split-ubiquitin system transformed the scFv-GPCR interaction signal into the expression of reporter genes. By optimizing the topological structure of scFv fusion protein and key elements, including signal peptides, transmembrane peptides, and flexible linkers, a system named Antigen-Antibody Co-Display (AACD) was established, which rapidly detected the interactions between antibodies and their target GPCRs, CXCR4 and CXCR5, while also determining the off-target antibodies and antibody-associated epitopes. The AACD system can rapidly determine the association between GPCRs and their candidate antibodies and shorten the research period for off-target detection and epitope identification. This system should improve the process of GPCR antibody development and provide a new strategy for GPCRs antibody screening.
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DOI：
10.1126/science.1194396
发表时间：
2010-11-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Wu B;Chien EY;Mol CD;Fenalti G;Liu W;Katritch V;Abagyan R;Brooun A;Wells P;Bi FC;Hamel DJ;Kuhn P;Handel TM;Cherezov V;Stevens RC
通讯作者：
Stevens RC
DOI：
10.1038/nri2744
发表时间：
2010-05
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
通讯作者：
--
影响因子：
1.6
作者：
Ohtake, Satoshi;Kita, Yoshiko;Arakawa, Tsutomu
通讯作者：
Arakawa, Tsutomu
影响因子：
5.3
作者：
Kaplon, Helene;Muralidharan, Mrinalini;Reichert, Janice M.
通讯作者：
Reichert, Janice M.
影响因子：
4.6
作者：
Li J;Gao J;Han L;Zhang Y;Guan W;Zhou L;Yu Y;Han W
通讯作者：
Han W