Abstract

Trans-2-nonenal is a fatty aldehyde with a long-chain, containing nine carbons. It was known that trans-2-nonenal is a cause of the odor associated with aging, and mainly found from the ages of 40 years. Phage display was a famous laboratory technique for proteins, peptides, and DNA interaction using bacteriophages to connect proteins with the genetic information. In this study, we have carried out random phage display screening to identify specific binding peptides for trans-2-nonenal. Trans-2-nonenal was directly coated on the plate. To select the phages against trans-2-nonenal, the random phages were attached to trans-2-nonenal and selected by 1–5 panning steps. Individual DNA of phage after 5 panning was analyzed to find the specific peptide sequence against trans-2-nonenal. The affinity of positive and negative binding phages was verified by ELISA assay using M13 phage-specific antibody. Among the phages, the phage with sequence of AHKSKLHQHVMFGGG (called as P4) in the end of tail, has shown the highest response. To explore a role of the peptide selected in sequence analysis and ELISA assay, the peptide was connected to magnetic beads. The peptide-coated beads were treated within trans-2-nonenal: treatment of P4 peptide shows significant decrease of trans-2-nonenal compared to negative peptide. Based on our results, it is suggested that the peptide, which is selected by phage display, could be used for the removal of trans-2-nonenal and odor associated with aging.

中文翻译：

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通过噬菌体展示筛选的特定的非核结合肽P4可以去除反式-2-非核

摘要

反式-2-壬烯醛是具有9个碳原子的长链脂肪醛。众所周知，反式-2-壬烯醛是与衰老相关的气味的原因，并且主要发现于40岁。噬菌体展示是一种著名的实验室技术，它利用噬菌体将蛋白质与遗传信息联系起来，从而进行蛋白质，肽和DNA的相互作用。在这项研究中，我们进行了随机噬菌体展示筛选，以鉴定反式-2-壬烯醛的特异性结合肽。将反式-2-壬烯醛直接包被在板上。为了选择针对反式-2-壬烯醛的噬菌体，将随机的噬菌体附着于反式-2-壬烯醛，并通过1-5个淘选步骤进行选择。分析5次淘选后的噬菌体的单个DNA，以发现针对反式-2-壬烯醛的特异性肽序列。使用M13噬菌体特异性抗体通过ELISA分析验证阳性和阴性结合噬菌体的亲和力。在噬菌体中，在尾巴末端具有AHKSKLHQHVMFGGG序列的噬菌体（称为P4）显示出最高的响应。为了探索所选肽在序列分析和ELISA分析中的作用，将该肽连接到磁珠上。肽包被的珠在反式2-壬烯醛内处理：与阴性肽相比，P4肽的处理显示反式-2-壬烯醛显着减少。根据我们的结果，建议通过噬菌体展示选择的肽可用于去除反式-2-壬烯醛和与衰老相关的气味。表现出最高的反应。为了探索所选肽在序列分析和ELISA分析中的作用，将该肽连接到磁珠上。肽包被的珠子在反式2-壬烯醛内处理：与阴性肽相比，P4肽的处理显示反式-2-壬烯醛显着减少。根据我们的结果，建议通过噬菌体展示选择的肽可用于去除反式-2-壬烯醛和与衰老相关的气味。表现出最高的反应。为了探索所选肽在序列分析和ELISA分析中的作用，将该肽连接到磁珠上。肽包被的珠子在反式2-壬烯醛内处理：与阴性肽相比，P4肽的处理显示反式-2-壬烯醛显着减少。根据我们的结果，建议通过噬菌体展示选择的肽可用于去除反式-2-壬烯醛和与衰老相关的气味。