Pathogen-specific deep sequence-coupled biopanning: A method for surveying human antibody responses.
Pathogen-specific deep sequence-coupled biopanning: A method for surveying human antibody responses.
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DOI：
10.1371/journal.pone.0171511
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Peabody DS
中科院分区：
文献类型：
--
作者：
Frietze KM;Pascale JM;Moreno B;Chackerian B;Peabody DS
在感染期间确定抗体反应的靶点对于设计疫苗、开发诊断和预后工具以及了解发病机制具有重要意义。我们开发了一种新的深度序列偶联生物筛选方法，能够识别存在于人多克隆血清中的抗体的蛋白质表位。在这里，我们报道了这种方法在鉴定急性感染期间引发的抗体识别的病原体特异性表位方面的适应性。作为一项原理验证，我们将这种方法应用于评估登革热病毒（DENV）抗体。利用急性继发性DENV感染患者的血清，我们筛选了一个显示在噬菌体MS2病毒样颗粒表面的DENV抗原片段文库，并通过深度序列分析表征了亲和选择的肽表位群体。尽管个体的反应存在相当大的差异，但我们发现包膜糖蛋白和非结构蛋白1中的几个表位普遍富集。该报告建立了一种表征人类血清中病原体特异性抗体反应的新方法，并在确定新的诊断和疫苗靶点方面具有未来的实用性。
Identifying the targets of antibody responses during infection is important for designing vaccines, developing diagnostic and prognostic tools, and understanding pathogenesis. We developed a novel deep sequence-coupled biopanning approach capable of identifying the protein epitopes of antibodies present in human polyclonal serum. Here, we report the adaptation of this approach for the identification of pathogen-specific epitopes recognized by antibodies elicited during acute infection. As a proof-of-principle, we applied this approach to assessing antibodies to Dengue virus (DENV). Using a panel of sera from patients with acute secondary DENV infection, we panned a DENV antigen fragment library displayed on the surface of bacteriophage MS2 virus-like particles and characterized the population of affinity-selected peptide epitopes by deep sequence analysis. Although there was considerable variation in the responses of individuals, we found several epitopes within the Envelope glycoprotein and Non-Structural Protein 1 that were commonly enriched. This report establishes a novel approach for characterizing pathogen-specific antibody responses in human sera, and has future utility in identifying novel diagnostic and vaccine targets.