文献类型：期刊文献

英文题名：Open-vessel polymerization of N-carboxyanhydride (NCA) for polypeptide synthesis

作者：Wu, Yueming[1,2];Chen, Kang[2];Wang, Jiangzhou[2];Dai, Wenhui[2];Yu, Haowen[2];Xie, Xinyi[2];Chen, Minzhang[2];Liu, Runhui[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Frontiers Sci Ctr Materiobiol & Dynam Chem, Engn Res Ctr Biomed Mat, Sch Mat Sci & Engn,Minist Educ,Key Lab Ultrafine M, Shanghai 200237, Peoples R China;[3]Chinese Acad Sci, Suzhou Inst Biomed Engn & Technol, Dept Biomat & Stem Cells, Suzhou, Peoples R China

年份：2025

卷号：20

期号：3

外文期刊名：NATURE PROTOCOLS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001331238200001)】；

基金：This research was supported by the National Natural Science Foundation of China (Nos. T2325010, 52203162 and 22075078), the National Key Research and Development Program of China (2022YFC2303100), Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism (Shanghai Municipal Education Commission), the Fundamental Research Funds for the Central Universities (JKVD1241029 and JKD01241701). The authors also thank the Research Center of Analysis and Test of East China University of Science and Technology for help on the characterization of studies reviewed in this paper.

语种：英文

摘要：Synthetic polypeptides, also known as poly(alpha-amino acids), have the same polyamide backbone structures as natural proteins and peptides. As an important class of biomaterials, polypeptides have been widely used because of their biocompatibility, bioactivity and biodegradability. Ring-opening polymerization of N-carboxyanhydride (NCA) is a classical and widely used method for the synthesis of polypeptides. The dominantly used primary amine-initiated NCA polymerization can yield well-defined polymers and complex macromolecular architectures, but the reaction is slow and sensitive to moisture, making it necessary to use anhydrous solvents and a glovebox. One solution is to use lithium hexamethyldisilazide (LiHMDS) as the initiator, as described in this protocol. LiHMDS-initiated NCA polymerization is less sensitive to moisture and can be carried out in an open vessel outside the glovebox. It is also very fast; the reaction can be complete within 5 min to produce 30-mer polypeptides. In this protocol, poly(gamma-benzyl-l-glutamate) is prepared as an example, but the protocol can easily be adapted to the synthesis of other polypeptides by generating NCAs from different amino acids, making it particularly suitable for the efficient parallel synthesis of polypeptide libraries. We provide detailed procedures for NCA synthesis and purification, the method of polymer end-group modification and measurement of polymerization kinetics and reactivity ratio. The procedure for synthesis of monomers and polymerization to form polypeptides requires <1 d. The superfast and open-vessel NCA polymerization method described here will probably enable a wide range of applications in the synthesis and functional study of polypeptide biomaterials.