毛细管电泳质谱联用技术测定肽和蛋白质混合物  

On line Capillary Electrophoresis mass Spectrometry for Determination of Peptide Mixture and Protein Mixture

作　　者：周国华 罗国安[1,2,3] 周克玉 古卓良[1,2,3] 

机构地区：[1]清华大学生命科学与工程研究院 [2]中国科学院南京土壤研究所 [3]南京军区后勤部药品检验所

出　　处：《高等学校化学学报》1998年第3期353-358,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学重点基金

摘　　要：以血管紧张素Ⅰ、Ⅱ、Ⅲ、Ｐ－物质和生长激素抑制剂混合物为测定对象，研究了未涂层柱、线性聚丙烯酰胺（ＬＰＡ）涂层柱和胺涂层柱对毛细管电泳分离肽类混合物的影响．结果表明：适合质谱测定的缓冲液不能在未涂层柱上有效地分离５种肽类混合物，而ｐＨ为５．０和４．５的ＮＨ４Ａｃ缓冲液却能在ＬＰＡ涂层柱和胺涂层柱上很好地分离上述物质．用ＣＥ－ＥＳＩ－ＭＳ法分离鉴定了５种生物活性肽和蛋白质混合物中的肌红蛋白、碳酸酐酶Ⅱ，实测了促红细胞生成素（ＥＰＯ）的胰酶消解肽图，并对ＣＥ－ＭＳ联用测定中的影响因素进行了讨论．The influence of uncoated, coated linear polyacrylamide(LPA) and amino coated capillaries on the separation of five peptide mixtures, including angiotensin Ⅰ, Ⅱ, Ⅲ, substance P and somatostain, was studied. The results showed that the peptide mixtures could not be separated completely in uncoated capillary by volatile buffer which was suitable for the MS analysis, whereas they could be separated in LPA and amino coated capillary by pH 5.0 and pH 4.5 NH 4Ac buffer respectively. In addition, the above peptide mixtures, protein mixtures comprised of myoglobin and carbonic anhydrase Ⅱ, tryptic digests of erythropoietin, were separated and characterized by capillary electrophoresis mass spectrometry(CE ESI MS) with electrospray ionization interface. The factors affecting the determination in CE ESI MS were also discussed.

关 键 词：肽 蛋白质 EPO胰酶肽图 CE-MS 血管紧张素 

分 类 号：Q510.3[生物学—生物化学] Q514.303