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刊物信息

期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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液质联用对重组胰高血糖素样肽-1-Fc融合蛋白中的制品相关蛋白进行定性定量分析

Identification and quantification of the product-related proteins in recombinant GLP-1-Fc fusion protein using UPLC-MS

作者(英文):
分类号:R917
出版年·卷·期(页码):2018,38 (11):1899-1904
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的:对重组胰高血糖素样肽-1-Fc融合蛋白(rGLP-1-Fc)的制品相关蛋白进行定性及定量分析。方法:利用液质联用法,对rGLP-1-Fc原液中制品相关蛋白的修饰类型进行鉴定;通过胰蛋白酶酶切液质肽图的方法,对修饰位点进行鉴定;采用反相超高效液相色谱法,测定3批原液中制品相关蛋白的比例。结果:该蛋白原液中主要存在3种制品相关蛋白;液质联用检测结果显示:1种为氧化及糖化的修饰产物,另外2种为一条二硫键解链导致的变异体;液质肽图分析发现氧化及糖化发生在第28位赖氨酸残基;对3批原液进行了检测,3种制品相关蛋白的平均含量分别为(2.37±0.04)%、(9.07±0.20)%和(4.03±0.07)%。结论:利用液质联用技术对rGLP-1-Fc原液中的制品相关蛋白进行了初步的定性、定量分析,测定结果可为同类产品的制品相关蛋白分析提供技术支持。

-----英文摘要:---------------------------------------------------------------------------------------

Objective: To identify and quantify the product-related proteins in the bulk of recombinant GLP-1-Fc fusion protein (rGLP-1-Fc). Methods: Modifications of the product-related proteins were identified using ultra performance liquid chromatography-mass spectrometry (UPLC-MS). The modified site was determined by trypsin-digested peptide LC-MS mapping. The percentages of product-related proteins in three batches of bulk were detected by reversed-phase UPLC. Results: Three types of product-related proteins were found in the bulk,and results of UPLC-MS showed that one was oxidized and glycated protein and the other two were the variants caused by breaking of one disulfide bond. The modification site of oxidation and glycation was K28 determined by peptide mass mapping. The average percentage contents of the three product-related proteins in three batches of bulk were (2.37±0.04)%,(9.07±0.20)% and (4.03±0.07)%,respectively. Conclusion: The product-related proteins in rGLP-1-Fc were identified and quantified preliminarily by UPLC-MS,which could provide technical support for product-related protein analysis of similar products.

-----参考文献:---------------------------------------------------------------------------------------
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