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一种用于高通量肽生产的快速手动固相肽合成方法。

A Rapid Manual Solid Phase Peptide Synthesis Method for High-Throughput Peptide Production.

作者信息

Overby Clyde, Abraham Brittany, Adjei-Sowah Emmanuella, March Alyson, Ling Kevin, Basu Sayantani, Benoit Danielle S W

机构信息

Department of Biomedical Engineering, University of Rochester, Rochester, New York, USA.

Center for Musculoskeletal Research, University of Rochester, Rochester, New York, USA.

出版信息

J Biomed Mater Res A. 2025 May;113(5):e37922. doi: 10.1002/jbm.a.37922.

Abstract

Solid phase peptide synthesis (SPPS) techniques are critical for developing and using peptides in various biomedical applications. However, typical synthesis routes used in SPPS are either resource-intensive (e.g., with automated synthesis or commercial services) or time-consuming (e.g., with manual benchtop synthesis). Here, a rapid manual synthesis method was developed to produce up to 8 peptides with fast cycle times simultaneously. Peptides synthesized manually were of equivalent or superior quality to those produced by in-house microwave-assisted automated peptide synthesis, with higher average crude purity of 70% compared to 50%. The method significantly reduced synthesis time, enabling the parallel coupling of up to 8 amino acids simultaneously in 15-20 min, as opposed to traditional benchtop peptide synthesis, which requires 80-150 min per amino acid. This approach offers an intermediate throughput between milligram-scale libraries and gram-scale single peptide synthesis, enabling rapid iteration for novel peptide designs without the need for expensive automated systems. As a result, peptide modifications, including incorporation of unnatural amino acids, can be explored, accelerating the development of peptides for a wide range of applications.

摘要

固相肽合成(SPPS)技术对于在各种生物医学应用中开发和使用肽至关重要。然而,SPPS中使用的典型合成路线要么资源密集型(例如,通过自动合成或商业服务),要么耗时(例如,通过手动台式合成)。在此,开发了一种快速手动合成方法,可同时快速循环生产多达8种肽。手动合成的肽与内部微波辅助自动肽合成产生的肽质量相当或更优,平均粗纯度更高,分别为70%和50%。该方法显著缩短了合成时间,与传统台式肽合成相比,能够在15至20分钟内同时平行偶联多达8种氨基酸,传统台式肽合成每添加一种氨基酸需要80至150分钟。这种方法提供了介于毫克级文库和克级单肽合成之间的中等通量,无需昂贵的自动化系统即可实现新型肽设计的快速迭代。因此,可以探索肽修饰,包括掺入非天然氨基酸,从而加速用于广泛应用的肽的开发。

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