基于2020年版《中华人民共和国药典》相关蛋白检测方法并联合质谱分析人干扰素α2b相关蛋白的一级结构  

Analysis the primary structure of human interferon α2b related proteins based on 2020《Pharmacopoeia of the People’s Republic of China》 related proteins detection method combined mass spectrometry

作　　者：朱秋媚 刘玉林 俞露 王江林 梁冠桥 孙瑞欣 刘琳琳 刘景会 ZHU Qiu-mei;LIU Yu-lin;YU Lu;WANG Jiang-lin;LIANG Guan-qiao;SUN Rui-xin;LIU Lin-lin;LIU Jing-hui(Changchun Institute of Biological Prodlucts Co.,Ltd.,Changehun 130012,China)

机构地区：[1]长春生物制品研究所有限责任公司,长春130012

出　　处：《药物分析杂志》2022年第6期1096-1100,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:用人干扰素α2b相关蛋白检测方法联合质谱对人干扰素α2b相关蛋白进行一级结构分析,确认不同保留时间的相关蛋白。方法:液相色谱条件为Agilent ZORBAX C_(18)色谱柱(250 mm×4.6 mm,300?,5μm);流动相A为30%乙腈水溶液(含0.2%三氟乙酸),流动相B为80%乙腈水溶液(含0.2%三氟乙酸),流速为0.5 mL·min^(-1),柱温为室温。质谱条件为电喷雾正离子模式,采用MS模式进行扫描,毛细管电压为2500 V,Cone电压为40 V,去溶剂气体温度为350℃,源温为120℃,去溶剂气体流速为800 L·h^(-1),扫描范围为m/z 400~4000,碰撞电压为40 V,MS采集时间为20~60 min。结果:相关蛋白的相对分子质量均与理论相对分子质量一致,3号峰为主峰,相对分子质量为19264.81和19395.93,为人干扰素α2b和N-末端甲硫氨酸化人干扰素α2b;1号峰相对分子质量为19280.72,为氧化人干扰素α2b,2号峰相对分子质量为19412.25,为(氧化+N-末端甲硫氨酸化)人干扰素α2b;4号峰相对分子质量为19265.13,为人干扰素α2b;5号峰相对分子质量为19322.81,为(氧化+N-末端乙酰化)人干扰素α2b;6号峰相对分子质量为19306.79,为N-末端乙酰化人干扰素α2b。结论:确认主峰为人干扰素α2b和N-末端甲硫氨酸化的人干扰素α2b,确认保留时间在主峰前面的相关蛋白分别为氧化和(氧化+N-末端甲硫氨酸化)修饰的人干扰素α2b,保留时间在主峰后面的相关蛋白分别为(氧化+N-末端乙酰化)和N-末端乙酰化修饰的人干扰素α2b。Objective:To analyze the primary structure of human interferonα2 b related proteins by using human interferonα2 b related proteins detection method combined with mass spectrometry,and to identify related proteins with different retention times.Methods:Agilent ZORBAX C_(18)(250 mm×4.6 mm,300?,5μm)column wasused and mobile phase A was 30%acetonitrile(0.2%TFA),mobile phase B was 80%acetonitrile(0.2%TFA).The flow rate was 0.5 mL·min^(-1),column temperature was room temperature.Electrospray positive ion mode with MS mode scaning was used.Capillary voltage was 2500 V,cone voltage was 40 V,desolvation temperature was 350℃,source temperature was 120℃,desolvation gas flow was 800 L·h^(-1),scan scope was 400-4000 m/z,collision voltage was 40 V and MS acquisition time was 20-60 min.Results:The molecular weight of the related proteins was consistent with the theoretical relative molecular mass.Peak 3 was the main peak,which relative molecular mass were 19264.81 and 19395.93.This peak was human interferonα2 b and N-terminal methionine human interferonα2 b.The peak 1 with relative molecular mass of 19280.72,was oxidized human interferonα2 b;and the peak 2 with relative molecular mass of 19412.25 was(oxidation+N-terminal methioninylation)human interferonα2 b.Peak 4 was human interferonα2 b with relative molecular mass at 19265.13.Peak 5 was(oxidation+N-terminal acetylation)human interferonα2 b with the relative molecular mass of 19322.81.Peak 6 was N-terminal acetylation of human interferonα2 b with the relative molecular mass of 19306.79.Conclusions:The main peak are human interferonα2 b and N-terminal methionine human interferonα2 b and the related proteins with retention time in front of the main peak were identified as oxidized and(oxidized+N-terminal methioninylation)modified human interferonα2 b.The related proteins with retention time behind the main peak were(oxidation+N-terminal acetylation)and N-acetylated modified human interferonα2 b.

关 键 词：人干扰素α2b 相关蛋白 一级结构 N-末端甲硫氨酸化 氧化 N-末端乙酰化 

分 类 号：R917[医药卫生—药物分析学]