| 液质联用研究人重组干细胞因子一级结构 |
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| 引用本文： | 郑建华,张平,饶晓璐,王世媛,颜江华.液质联用研究人重组干细胞因子一级结构[J].厦门大学学报(自然科学版),2011,50(3):612-616. |
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| 作者姓名： | 郑建华 张平 饶晓璐 王世媛 颜江华 |
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| 作者单位： | 1. 厦门大学医学院,福建厦门,361005
2. 厦门特宝生物工程股份有限公司,福建厦门,361028 |
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| 摘 要： | 通过高效液相色谱(HPLC)和质谱技术,对人重组干细胞因子(rhSCF)分子进行分析,以确证其一级结构.将rh-SCF样品胰酶酶切,HPLC-RPC分离,基质辅助激光解吸附电离-飞行时间串联质谱仪(MALDI-TOF-MS)对rhSCF的酶切肽段进行解析,并对rhSCF的O-糖基化和N-糖基化位点进行了分析.结果:(1)rhSCF分子质量得到测定;(2)rhSCF的一级结构得到确认,测定的肽质量谱的覆盖率达到94%,分子中有两对二硫键;(3)鉴定了样品中的N-糖基化位点,并对rhSCF产品的O-及N-连接糖型上的糖结构进行初步探讨.结果显示:rhSCF一级氨基酸序列和二硫键形成位置与理论上一致,糖基化特性符合毕赤酵母表达蛋白的糖基化特点.
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| 关 键 词： | MALDI-TOF-MS 人重组干细胞因子 二硫键 糖基化 |
Primary Structure Determination of Recombinant Human Stem Cell Factor by HPLC and MALDI-TOF |
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| ZHENG Jian-hua,ZHANG Ping,RAO Xiao-lu,WANG Shi-yuan,YAN Jiang-hua.Primary Structure Determination of Recombinant Human Stem Cell Factor by HPLC and MALDI-TOF[J].Journal of Xiamen University(Natural Science),2011,50(3):612-616. |
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| Authors: | ZHENG Jian-hua ZHANG Ping RAO Xiao-lu WANG Shi-yuan YAN Jiang-hua |
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| Institution: | ZHENG Jian-hua1,ZHANG Ping2,RAO Xiao-lu2,WANG Shi-yuan2,YAN Jiang-hua1(1.Medical College of Xiamen University,Xiamen 361005,China,2.Xiamen Amoytop Biotech Corporation Limited,Xiamen 361028,China) |
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| Abstract: | High performance liquid chromatography(HPLC)and matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS)were used to determine the primary structure of the recombinant human stem cell factor(rhSCF) samples.The rhSCF samples were digested by TPCK-trypsin,and separated by HPLC.The molecular weight of rhSCF and the amino sequences of its the peptides were determined by MALDI-TOF,and the glycosylated peptides were analyzed.The results are as follows:(1) the mass of rhSCF was de... |
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| Keywords: | MALDI-TOF-MS recombinant human stem cell factor disulfide bond oligosaccharide |
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