Identification of disease-specific motifs in the antibody specificity repertoire via next-generation sequencing.
Identification of disease-specific motifs in the antibody specificity repertoire via next-generation sequencing.
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DOI：
10.1038/srep30312
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发表时间：
2016-08-02
影响因子：
4.6
通讯作者：
Daugherty PS
中科院分区：
文献类型：
--
作者：
Pantazes RJ;Reifert J;Bozekowski J;Ibsen KN;Murray JA;Daugherty PS
疾病特异性抗体可以作为高效的生物标志物，但仅被鉴定用于相对少数的自身免疫性疾病。开发了一种通过整合细菌展示肽库筛选、下一代测序（NGS）和计算分析来识别疾病特异性结合基序的方法。通过使用细胞分选和进行 NGS 识别每个样本的结合肽库成员来确定抗体特异性库。开发并应用了一种称为使用下一代测序实验识别基序（IMUNE）的计算算法来发现疾病和健康对照特异性基序。 IMUNE 执行全面的模式搜索，识别疾病组或对照组中统计上丰富的模式，并对模式进行聚类以生成主题。使用乳糜泻血清作为发现集，除了新的基序之外，IMUNE 在验证血清集中鉴定了一个具有高诊断敏感性和特异性的共有基序 (QPEQPF[PS]E)。因此，肽展示和测序 (Display-Seq) 与 IMUNE 分析相结合可能有助于表征抗体库并鉴定疾病特异性抗体表位和生物标志物。
Disease-specific antibodies can serve as highly effective biomarkers but have been identified for only a relatively small number of autoimmune diseases. A method was developed to identify disease-specific binding motifs through integration of bacterial display peptide library screening, next-generation sequencing (NGS) and computational analysis. Antibody specificity repertoires were determined by identifying bound peptide library members for each specimen using cell sorting and performing NGS. A computational algorithm, termed Identifying Motifs Using Next- generation sequencing Experiments (IMUNE), was developed and applied to discover disease- and healthy control-specific motifs. IMUNE performs comprehensive pattern searches, identifies patterns statistically enriched in the disease or control groups and clusters the patterns to generate motifs. Using celiac disease sera as a discovery set, IMUNE identified a consensus motif (QPEQPF[PS]E) with high diagnostic sensitivity and specificity in a validation sera set, in addition to novel motifs. Peptide display and sequencing (Display-Seq) coupled with IMUNE analysis may thus be useful to characterize antibody repertoires and identify disease-specific antibody epitopes and biomarkers.