摘要
目的:多肽与小分子化学药物相比,具有生物活性高、特异性强、不容易产生耐药性等特点,是目前新型药物研发的重点领域。多肽的合成直接影响到多肽药物的作用机制以及药物效果,因此需要建立一种更加便捷、高效的多肽合成方法。方法:采用Fmoc固相合成法合成多肽HF01,通过比较氨基酸连接的反应体系以及氨基酸脱保护的反应体系,从中确定最优体系。利用乙酰化基团进行肽链末端保护,经肽链剪切制备干燥的粗肽,最后采用高效液相色谱仪与高分辨质谱仪联用对粗肽进行纯化。结果:确定多肽合成的连接和脱保护反应体系,并获得纯度高达98.3%的线性多肽。结论:建立了一种高效、便捷的多肽合成及纯化方法,提高了实验室合成多肽的效率,为多肽类药物的研发提供技术支撑。
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.
作者
王宇航
陈学明
刘俗生
阮志军
张敏
宋春丽
尹丰
李子刚
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)
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2023年第1期35-41,共7页
China Biotechnology
关键词
多肽
Fmoc固相合成
氨基酸连接
液相色谱串联质谱
纯化
Polypeptide
Fmoc solid-phase synthesis
Amino acid connection
Liquid chromatography tandem mass spectrometry(LC-MS/MS)
Purification
