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Manufacturing method of stable isotope-labeled target peptide fragments in mass spectrometry
Manufacturing method of stable isotope-labeled target peptide fragments in mass spectrometry
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机译:质谱中稳定同位素标记的靶肽片段的制备方法
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摘要
An object of the present invention is to provide a method for producing a stable isotope-labeled target peptide fragment in mass spectrometry, which achieves inexpensive and convenient production. As a solution to attain the object, the stable isotope-labeled target peptide fragment in mass spectrometry is produced using a method comprising the steps of: expressing a DNA conjugate in a system having a stable isotope-labeled amino acid to thereby prepare a stable isotope-labeled protein, wherein the DNA conjugate comprises: a tandemly linked DNA in which two or more DNAs encoding one or more types of target peptide fragments are linked in tandem; and a DNA encoding a peptide fragment for concentration measurement; subjecting the stable isotope-labeled protein to fragmentation treatment with trypsin to prepare a stable isotope-labeled peptide fragment for concentration measurement and stable isotope-labeled target peptide fragments; quantifying the stable isotope-labeled peptide fragment for concentration measurement using a liquid chromatograph-tandem mass spectrometer (LC/MS/MS); and calculating the concentration of the stable isotope-labeled target peptide fragment of each type from the quantification value of the stable isotope-labeled peptide fragment for concentration measurement.
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机译:本发明的目的是提供一种在质谱中生产稳定的同位素标记的靶肽片段的方法,该方法实现了廉价且方便的生产。作为实现该目的的解决方案,使用以下方法来制备质谱中的稳定同位素标记的靶肽片段:在具有稳定同位素标记的氨基酸的系统中表达DNA缀合物,从而制备稳定同位素。 -标记的蛋白质,其中所述DNA缀合物包含:串联连接的DNA,其中两个或更多个编码一种或多种类型的目标肽片段的DNA串联连接;和编码用于浓度测量的肽片段的DNA;用胰蛋白酶对稳定的同位素标记的蛋白质进行片段化处理,以制备用于浓度测量的稳定的同位素标记的肽片段和稳定的同位素标记的靶肽片段;使用液相色谱-串联质谱仪(LC / MS / MS)定量稳定同位素标记的肽片段以进行浓度测量;根据浓度测定用的同位素标记肽片段的定量值,算出各类型的同位素标记靶肽片段的浓度。
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