Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands.
Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands.
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DOI：
10.1371/journal.pone.0160940
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Martinez JS
中科院分区：
文献类型：
--
作者：
Phipps ML;Lillo AM;Shou Y;Schmidt EN;Paavola CD;Naranjo L;Bemdich S;Swanson BI;Bradbury AR;Martinez JS
多肽是显微镜、生物传感和靶向递送的重要亲和配体。然而，因为它们对目标的亲和力很低，所以从大型的幼稚库中进行选择可能是具有挑战性的。当从展示文库中选择多肽配体时，重要的是：1)确保有效的展示；2)最大化选择高亲和力配体的能力；以及3)最小化展示上下文对结合的影响。“辅助细胞”包装系统被描述为一种工具，用于生产基于具有不同展示水平的噬菌体结构的丝状噬菌体颗粒，同时保持不受辅助噬菌体污染。在这里，我们报道了该系统首次用于多肽展示，包括辅助细胞的系统表征和优化，它们在抗体展示中的低效使用，以及它们在创建和选择一组噬菌体展示多肽库中的应用。我们的文库通过标准或深度测序以前所未有的精度进行了分析，并显示出比商业黄金标准更高的质量。利用我们的辅助细胞库，我们获得了识别鼠疫耶尔森菌表面抗原F1V和L谷氨酰胺结合周质蛋白QBP的配体。在后一种情况下，与迄今描述的任何肽库选择不同，我们使用噬菌体和酵母展示的组合来选择耐人寻味的多肽配体。基于我们的选择的成功，我们认为用辅助细胞获得的多肽库不仅适合，而且比传统的噬菌体展示文库更适合于多肽配体的选择。
Peptides are important affinity ligands for microscopy, biosensing, and targeted delivery. However, because they can have low affinity for their targets, their selection from large naïve libraries can be challenging. When selecting peptidic ligands from display libraries, it is important to: 1) ensure efficient display; 2) maximize the ability to select high affinity ligands; and 3) minimize the effect of the display context on binding. The “helper cell” packaging system has been described as a tool to produce filamentous phage particles based on phagemid constructs with varying display levels, while remaining free of helper phage contamination. Here we report on the first use of this system for peptide display, including the systematic characterization and optimization of helper cells, their inefficient use in antibody display and their use in creating and selecting from a set of phage display peptide libraries. Our libraries were analyzed with unprecedented precision by standard or deep sequencing, and shown to be superior in quality than commercial gold standards. Using our helper cell libraries, we have obtained ligands recognizing Yersinia pestis surface antigen F1V and L-glutamine-binding periplasmic protein QBP. In the latter case, unlike any of the peptide library selections described so far, we used a combination of phage and yeast display to select intriguing peptide ligands. Based on the success of our selections we believe that peptide libraries obtained with helper cells are not only suitable, but preferable to traditional phage display libraries for selection of peptidic ligands.
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影响因子：
36.8
作者：
BRUBAKER, RR
通讯作者：
BRUBAKER, RR
影响因子：
3.5
作者：
ROBERTS, D;GUEGLER, K;WINTER, J
通讯作者：
WINTER, J
DOI：
10.1084/jem.179.1.323
发表时间：
1994-01-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Rock EP;Sibbald PR;Davis MM;Chien YH
通讯作者：
Chien YH
DOI：
10.1038/nsb0298-140
发表时间：
1998-02-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Lubkowski, J;Hennecke, F;Wlodawer, A
通讯作者：
Wlodawer, A
影响因子：
11.4
作者：
FOLGORI, A;TAFI, R;NICOSIA, A
通讯作者：
NICOSIA, A