A two-step method preparation of semaglutide through solid-phase synthesis and inclusion body expression

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-03-23 DOI:10.1016/j.pep.2024.106477

Dezheng Peng , Yang Li , Linlin Si , Bo Zhu , Peng Wu , Yibang Li , Dongfang Tang , Yu Liu , Yunxiao Zhang

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Abstract

Semaglutide is currently the most promising antidiabetic drug, especially for the treatment of type 2 diabetes mellitus, due to its excellent efficacy in glycemic control and weight loss. However, the production of semaglutide remains high cost, and high yield, low cost, and high purity still remains a challenge. Herein, we reported a convenient and high-yield strategy for the preparation of semaglutide through fragmented condensation coupling, involving solid-phase peptide synthesis of tetrapeptide and on-column refolding and on-column enzyme cleavage based inclusion body expression of Lys²⁶Arg³⁴GLP-1 (11–37) with fused protein tags in an X-Y-D4K-G pattern. The optimized N-terminal protein tag significantly boosts inclusion body expression level, while on-column refolding and on-column enzyme cleavage avoid precipitation, enhancing efficiency and yield together with one-step purification. The successful preparation of semaglutide is expected to achieve large-scale industrial production with low cost, high yield and high purity.

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通过固相合成和包涵体表达两步法制备塞马鲁肽

由于在控制血糖和减轻体重方面的卓越疗效，塞马鲁肽是目前最有前途的抗糖尿病药物，尤其是用于治疗 2 型糖尿病。然而，塞马鲁肽的生产成本仍然很高，高产率、低成本和高纯度仍然是一个难题。在此，我们报道了一种通过片段缩合偶联制备塞马鲁肽的便捷、高产策略，包括四肽的固相肽合成、柱上重折叠以及基于包涵体表达的 X-Y-D4K-G 模式融合蛋白标签的 Lys26Arg34GLP-1 (11-37) 柱上酶切。优化的 N 端蛋白标签显著提高了包涵体的表达水平，柱上重折叠和柱上酶切避免了沉淀，一步纯化提高了效率和产量。塞马鲁肽的成功制备有望实现低成本、高收率、高纯度的大规模工业化生产。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.