Library Design-Facilitated High-Throughput Sequencing of Synthetic Peptide Libraries.
Library Design-Facilitated High-Throughput Sequencing of Synthetic Peptide Libraries.
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文库设计促进合成肽文库的高通量测序。
DOI：
10.1021/acscombsci.7b00109
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发表时间：
2017
影响因子：
--
通讯作者：
Pentelute,BradleyL
中科院分区：
文献类型：
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作者：
Vinogradov,AlexanderA;Gates,ZacharyP;Zhang,Chi;Quartararo,AnthonyJ;Halloran,KathrynH;Pentelute,BradleyL
报道了一种实现合成肽混合物的高通量从头测序的方法。该方法利用鸟枪纳米液体色谱法结合基于串联质谱的文库混合物（多达2000种肽）的从头测序以及自动化数据分析协议来过滤掉不正确的分配，噪音和合成副产物。为了增加测序结果的置信度，开发了质谱友好的文库设计，其能够每小时明确解码多达600个肽序列，同时在大多数情况下保持大于85%的序列识别率。通过匹配从文库测序样品中鉴定的选择的真实肽的片段化光谱，另外证实了报告的解码策略的可靠性。本文报道的方法直接适用于产生活性化合物混合物的筛选技术，包括单珠单化合物文库的颗粒分选和在溶液中进行的合成文库混合物的亲和富集。
A methodology to achieve high-throughput de novo sequencing of synthetic peptide mixtures is reported. The approach leverages shotgun nanoliquid chromatography coupled with tandem mass spectrometry-based de novo sequencing of library mixtures (up to 2000 peptides) as well as automated data analysis protocols to filter away incorrect assignments, noise, and synthetic side-products. For increasing the confidence in the sequencing results, mass spectrometry-friendly library designs were developed that enabled unambiguous decoding of up to 600 peptide sequences per hour while maintaining greater than 85% sequence identification rates in most cases. The reliability of the reported decoding strategy was additionally confirmed by matching fragmentation spectra for select authentic peptides identified from library sequencing samples. The methods reported here are directly applicable to screening techniques that yield mixtures of active compounds, including particle sorting of one-bead one-compound libraries and affinity enrichment of synthetic library mixtures performed in solution.