Phage-DMS: A Comprehensive Method for Fine Mapping of Antibody Epitopes.
Phage-DMS: A Comprehensive Method for Fine Mapping of Antibody Epitopes.
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Phage-DMS：一种用于抗体表位精细定位的综合方法。
DOI：
10.1016/j.isci.2020.101622
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发表时间：
2020-10-23
期刊：
影响因子：
5.8
通讯作者：
Overbaugh J
中科院分区：
文献类型：
--
作者：
Garrett ME;Itell HL;Crawford KHD;Basom R;Bloom JD;Overbaugh J
了解抗体反应对于开发疫苗和基于抗体的疗法至关重要，并激发了最近分离抗体的新方法的发展。确定特异性或允许逃逸的抗体-抗原相互作用的方法尚未跟上。我们开发了噬菌体-DMS，一种结合两种强大方法的方法-噬菌体肽库的免疫沉淀和深度突变扫描(DMS) -实现抗体表位的高通量精细定位。例如，我们设计了编码HIV包膜所有可能的氨基酸变体的序列来创建噬菌体文库。使用噬菌体- dms，我们确定了用其他方法预测的四种已知线性表位的HIV单克隆抗体的逃逸位点。在某些情况下，噬菌体- dms的结果细化了表位，超出了先前研究中确定的结果。这种方法有潜力在一次实验中快速和全面地筛选许多抗体，以确定结合相互作用所必需的位点。噬菌体- dms结合了噬菌体展示技术和深度突变扫描技术，噬菌体- dms鉴定了导致HIV逃避Env抗体结合的单个突变，噬菌体- dms中的突变与平行方法的结果相关；基因组文库
Understanding the antibody response is critical to developing vaccine and antibody-based therapies and has inspired the recent development of new methods to isolate antibodies. Methods to define the antibody-antigen interactions that determine specificity or allow escape have not kept pace. We developed Phage-DMS, a method that combines two powerful approaches—immunoprecipitation of phage peptide libraries and deep mutational scanning (DMS)—to enable high-throughput fine mapping of antibody epitopes. As an example, we designed sequences encoding all possible amino acid variants of HIV Envelope to create phage libraries. Using Phage-DMS, we identified sites of escape predicted using other approaches for four well-characterized HIV monoclonal antibodies with known linear epitopes. In some cases, the results of Phage-DMS refined the epitope beyond what was determined in previous studies. This method has the potential to rapidly and comprehensively screen many antibodies in a single experiment to define sites essential for binding interactions. A high-throughput, comprehensive method to identify antibody epitopes is needed Phage-DMS combines phage display technology and deep mutational scanning Phage-DMS identified single mutations that lead to escape from HIV Env antibody binding Effect of mutations in Phage-DMS correlate with results using a parallel approach Virology; Genomic Library
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