Ling V, Guzzetta A W, Canova-Davis E, Stults J T, Hancock W S, Covey T R, Shushan B I
Genentech, Inc., South San Francisco, California 94080.
Anal Chem. 1991 Dec 15;63(24):2909-15. doi: 10.1021/ac00024a020.
A detailed tryptic map is presented for recombinant human tissue plasminogen activator (rt-PA). Electrospray ionization mass spectrometry is utilized as an on-line HPLC detector for tryptic mapping of this glycoprotein. The additional dimension provided by mass spectrometry gives considerably more detail about the complex tryptic map and significantly enhances the high-resolution chromatographic separation by distinguishing by mass any coeluting components. Through this improvement, the proline isomers of a tryptic peptide were observed eluting over a broad range of retention times. The glycopeptides of rt-PA are observed as well as any corresponding nonglycosylated peptides. In addition, the carbohydrate heterogeneity is readily observed, allowing analysis of the carbohydrate composition. The characteristic diagonal patterns formed by glycopeptides in a contour plot of the data allow rapid recognition of the glycopeptides.
本文给出了重组人组织型纤溶酶原激活剂（rt-PA）的详细胰蛋白酶图谱。电喷雾电离质谱被用作该糖蛋白胰蛋白酶图谱分析的在线高效液相色谱检测器。质谱提供的额外维度能给出关于复杂胰蛋白酶图谱的更多细节，并且通过按质量区分任何共洗脱成分，显著增强了高分辨率色谱分离效果。通过这一改进，观察到一种胰蛋白酶肽的脯氨酸异构体在很宽的保留时间范围内洗脱。rt-PA的糖肽以及任何相应的非糖基化肽都被观察到。此外，很容易观察到碳水化合物的异质性，从而能够分析碳水化合物的组成。数据等高线图中糖肽形成的特征对角线模式有助于快速识别糖肽。
