A Solid-phase Synthesis Method and Purification of Polypeptides

doi:10.13523/j.cb.2209002

China Biotechnology

2023, Vol. 43

Issue (1): 35-41 DOI: 10.13523/j.cb.2209002

A Solid-phase Synthesis Method and Purification of Polypeptides

WANG Yu-hang,CHEN Xue-ming,LIU Su-sheng,RUAN Zhi-jun,ZHANG Min,SONG Chun-li,YIN Feng^*(

),LI Zi-gang^*(

)

Shenzhen Bay Laboratory Pingshan Translational Medicine Center, Shenzhen 518118, China

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Abstract

Objective: Compared with small molecule chemical drugs, peptides have the advantages of high biological activity, strong specificity, and drug resistance, and thus, they become one of the priorities of new drug research and development. Peptide synthesis is the premise of the research and development of polypeptide drugs. Therefore, it is very important to establish a more convenient and efficient synthesis method. Methods: Fmoc solid phase synthesis was used for HF01 synthesis of polypeptides. First, the optimal system was determined by comparing the amino acid linked reaction system and the amino acid deprotection reaction system, and the acetylation group was applied for peptide N-Term. Then, the peptides were cut off from the resin. Finally, the obtained peptides were purified using both an HPLC and a mass spectrometer. Results: The ligation and deprotection reaction system for polypeptide synthesis was determined, and the polypeptide was successfully synthesized and purified with high purification efficiency of 98.3%. Conclusion: Above all, the operation is simple and affordable. This method is commonly used for polypeptide synthesis and provides feasibility for polypeptide drug structure studies. It is expected that this reliable synthesis method will provide help for more laboratory researchers.

Key words： Polypeptide Fmoc solid-phase synthesis Amino acid connection Liquid chromatography tandem mass spectrometry (LC-MS/MS) Purification

Received: 02 September 2022 Published: 14 February 2023

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	Yu-hang WANG
	Xue-ming CHEN
	Su-sheng LIU
	Zhi-jun RUAN
	Min ZHANG
	Chun-li SONG
	Feng YIN
	Zi-gang LI

Cite this article:

WANG Yu-hang, CHEN Xue-ming, LIU Su-sheng, RUAN Zhi-jun, ZHANG Min, SONG Chun-li, YIN Feng, LI Zi-gang. A Solid-phase Synthesis Method and Purification of Polypeptides. China Biotechnology, 2023, 43(1): 35-41.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2209002 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I1/35

Fig.1 Polypeptide synthesis route

Table 1 Comparison of the two amino acid linked reaction systems

Table 2 Comparison of the reaction systems for the different deprotection groups

Table 3 High-performance liquid phase and mass spectrometry detection data for the polypeptide HF01

Fig.2 HPLC detection results

Fig.3 Mass spectrometry detection results


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