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Method for automatically parsing stable isotope labeled sugar chain quantification mass spectrum data

A stable isotope and automatic analysis technology, applied in the field of protein analysis, can solve the problems of quantitative error, easy omission of low-abundance sugar component quantitative results, weak response of sugar chain mass spectrometry, etc., to achieve high quantitative accuracy and robustness, Good application prospect, convenient calculation effect

Active Publication Date: 2019-05-10
FUDAN UNIV
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Problems solved by technology

Due to the microscopic inhomogeneity of the sugar chain structure and the low glycosidic bond energy, the response of sugar chain mass spectrometry is weak. In the face of massive sugar chain quantitative mass spectrometry data, manual analysis is not only cumbersome and time-consuming, but also prone to quantitative errors, and it is easy to miss low abundance. Quantitative results for high-degree and low-ionization-efficiency sugar components
When the mass difference of the isotope-labeled label on the sugar chain is small, it is easy to affect the accuracy of quantification due to isotope overlap
In addition, as the molecular weight of sugar chains increases, the distribution of theoretical isotopic peaks of sugar chains also changes accordingly, and the position of the highest theoretical isotopic peak also changes. The traditional quantitative method using monoisotopic peak intensity is prone to errors

Method used

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  • Method for automatically parsing stable isotope labeled sugar chain quantification mass spectrum data
  • Method for automatically parsing stable isotope labeled sugar chain quantification mass spectrum data
  • Method for automatically parsing stable isotope labeled sugar chain quantification mass spectrum data

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Embodiment Construction

[0026] Asialofetuin (ASF) was divided into two parts, one part was added with 100 mM ammonium bicarbonate and PNGase F enzyme; the other part was added with 100 mM ammonium bicarbonate and PNGase F enzyme dissolved in oxygen 18 water, 37 After incubation at ℃ for 16 hours, 10K ultrafiltration, PGC desalination, and freeze-drying were performed respectively to obtain the sugar chains of ASF. The obtained sugar chains were mixed according to two ratios of 1:1 and 1:5, and then detected by MALDI mass spectrometry, and then the data was analyzed by gquant. When the m / z of the sugar chain is greater than 2250, gGuant shows advantages, such as image 3 For the sugar chains shown in , gQuant reported that the quantitative ratios of the two were 1.084 and 4.816, which were close to the theoretical values, while the quantitative results of hand-picked but calculated isotope peaks were 1.4649 and 10.607, respectively. The serious deviation of the second ratio is caused by the severe in...

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Abstract

The invention belongs to the technical field of protein analysis, and particularly relates to a method for automatically parsing stable isotope labeled sugar chain quantification mass spectrum data. According to the invention, a tool gQuant is utilized to correct mass of an isotope labeled sugar chain produced by an MALDI-MS so as to carry out accurate relative quantification; the tool sufficiently considers the characteristics of isotope labeled relative quantification data; isotopes are alternatively and accurately corrected; moreover, particularly, an algorithm for carrying out quantification on the basis of the highest theoretical isotope intensity peak is set by utilizing a theoretical isotope distribution mode and a mass spectrum fracturing property of the sugar chain; and the methodhas higher quantification accuracy and robustness for a high-molecular-weight and low-abundance sugar chain component.

Description

technical field [0001] The invention belongs to the technical field of protein analysis, and specifically relates to a method for accurate relative quantification by correcting the mass of isotope-labeled sugar chains produced by matrix-assisted laser desorption mass spectrometry (MALDI-MS). Background technique [0002] Kylation, as an important post-translational modification of proteins, is widely involved in many biological and physiological processes in life, including sperm-egg binding (cell recognition and interaction), signal transduction, protein folding, and protein half-life. At present, the isotope labeling quantitative technology based on mass spectrometry is one of the most widely used technologies in the quantitative means of sugar chains. Due to the microscopic inhomogeneity of the sugar chain structure and the low glycosidic bond energy, the response of sugar chain mass spectrometry is weak. In the face of massive sugar chain quantitative mass spectrometry d...

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N33/68
Inventor 黄江铭蒋碧云曹纬倩杨芃原
Owner FUDAN UNIV
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