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    METHOD FOR THE DETERMINATION OF SEQUENCE VARIANTS OF POLYPEPTIDES
    1.
    发明申请
    METHOD FOR THE DETERMINATION OF SEQUENCE VARIANTS OF POLYPEPTIDES 审中-公开
    用于确定多肽序列变异体的方法

    公开(公告)号：US20120322093A1

    公开(公告)日：2012-12-20

    申请号：US13579402

    申请日：2011-02-16

    申请人： Hans Koll ,  Jeorg Thomas Regula ,  Frank Wiegeshoff ,  Anne Zeck

    发明人： Hans Koll ,  Jeorg Thomas Regula ,  Frank Wiegeshoff ,  Anne Zeck

    IPC分类号： G01N27/62 ,  C12P21/00

    CPC分类号： C12Q1/37 ,  G01N33/6848 ,  G01N2560/00

    摘要： The current invention is directed to a method for determining amino acid sequence mutations in a produced polypeptide, comprising the following steps of a) providing a sample of a produced polypeptide, b) incubating the polypeptide in the sample with a protease, c) performing a two dimensional analysis using reversed phase chromatography coupled with a high resolution mass spectroscopy (FT-ICR/FT-orbitrap) and MS/MS analysis of the amino acid sequence fragments of the peptides, d) data evaluation by comparing the LC-MS data sets obtained for the samples side by side with the data set of a reference sample, by searching for differences in the signal intensities at given retention times and by evaluation of differential signals with respect to amino acid sequence mutations. The reference sample for data evaluation (d) can be either a well characterized standard or one of the samples to be analyzed.

    摘要翻译： 本发明涉及一种用于测定产生的多肽中的氨基酸序列突变的方法，包括以下步骤：a）提供产生的多肽的样品，b）将样品中的多肽与蛋白酶一起孵育，c） 使用反相色谱与高分辨率质谱（FT-ICR / FT-orbitrap）相结合的二维分析和肽的氨基酸序列片段的MS / MS分析，d）通过比较LC-MS数据集进行数据评估 通过在给定的保留时间上搜索信号强度的差异，并且通过评估关于氨基酸序列突变的差异信号，将样品与参考样品的数据集并排获得。 用于数据评估的参考样本（d）可以是一个良好表征的标准或要分析的样品之一。

    IMMUNOGLOBULIN GLYCOSYLATION PATTERN ANALYSIS
    2.
    发明申请
    IMMUNOGLOBULIN GLYCOSYLATION PATTERN ANALYSIS 审中-公开
    免疫球蛋白糖蛋白模式分析

    公开(公告)号：US20110294150A1

    公开(公告)日：2011-12-01

    申请号：US13147914

    申请日：2010-02-05

    申请人： Hans Koll ,  Joerg Thomas Regula ,  Peter Sondermann ,  Anne Zeck

    发明人： Hans Koll ,  Joerg Thomas Regula ,  Peter Sondermann ,  Anne Zeck

    IPC分类号： C12Q1/37 ,  C12P21/06

    CPC分类号： G01N33/6854 ,  C07K1/20 ,  C07K1/34 ,  C07K16/00 ,  C07K2317/24 ,  C07K2317/41 ,  C07K2317/50 ,  G01N33/6848 ,  G01N2400/00

    摘要： The current invention is directed to a method for the determination of the glycosylation pattern of a human immunoglobulin of the subclass IgG1 or IgG4 or of a murine immunoglobulin of the subclass Ig-G2a or IgG3 comprising the following steps: a) cleaving said immunoglobulin into fragments by enzymatic digestion with the enzyme IdeS, b) separating the fragments of said immunoglobulin obtained by the enzymatic digestion by reversed phase high performance liquid chromatography, c) subjecting the separated fragments of said immunoglobulin obtained in step b) to a mass spectrometric analysis, and d) determining the glycosylation pattern of said immunoglobulin from the mass spectrometric data obtained in step c).

    摘要翻译： 本发明涉及确定亚类IgG1或IgG4的人免疫球蛋白或IgA-G2a或IgG3亚类的鼠免疫球蛋白的糖基化模式的方法，包括以下步骤：a）将所述免疫球蛋白切割成片段 通过用酶IdeS进行酶消化，b）通过反相高效液相色谱法分离通过酶消化获得的所述免疫球蛋白的片段，c）将步骤b）中获得的所述免疫球蛋白分离的片段进行质谱分析，以及 d）从步骤c）中获得的质谱数据确定所述免疫球蛋白的糖基化模式。