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    • 1. 发明申请
    • METHODS OF ANALYZING AND PREPARING PROTEIN COMPOSITIONS
    • 分析和制备蛋白质组合物的方法
    • WO2013148323A1
    • 2013-10-03
    • PCT/US2013/032171
    • 2013-03-15
    • SHIRE HUMAN GENETIC THERAPIES, INC.LIN, MelanieSALINAS, PaulWU, Shiaw-lin
    • SHIRE HUMAN GENETIC THERAPIES, INC.LIN, MelanieSALINAS, PaulWU, Shiaw-lin
    • A61K38/46
    • C12Q1/44G01N2560/00
    • Methods for analyzing protein samples of recombinant human arylsulfatase A(rhASA), having one or more disulfide linkages are described. In some instances, the samples are analyzed using enzyme digestion, e.g., multi-enzyme digestion and liquid chromatography-mass spectrometry (LC-MS). The proteins described herein can be glycosylated (e.g., fully or partially glycosylated) or non-glycosylated. The methods described herein can include one or more of enzymatic digestion, chromatography, and mass spectrometry, e.g., multi-enzyme digestion and/or liquid chromatography-mass spectrometry (LC-MS). In some embodiments, the mass spectrometry utilizes collision induced dissociation (CID) and/or electron transfer dissociation (ETD). For example, the mass spectrometry can utilize CID, ETD, and CID of the isolated charge-reduced ions (MS3), e.g., after ETD.
    • 描述了分析具有一个或多个二硫键的重组人芳基硫酸酯酶A(rhASA)的蛋白质样品的方法。 在一些情况下,使用酶消化(例如多酶消化和液相色谱 - 质谱法(LC-MS))分析样品。 本文所述的蛋白质可以被糖基化(例如完全或部分糖基化)或非糖基化。 本文所述的方法可以包括酶消化,色谱法和质谱法中的一种或多种,​​例如多酶消化和/或液相色谱 - 质谱(LC-MS)。 在一些实施方案中,质谱法利用碰撞诱导解离(CID)和/或电子转移离解(ETD)。 例如,质谱可以利用分离的电荷还原离子(MS3)的CID,ETD和CID,例如在ETD之后。