N-sugar chain relative quantitation method based on 18O mark
A relatively quantitative, sugar chain technology used in analytical chemistry to solve problems such as isotopic peak overlap
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[0017] The specific steps of the present invention are further described below in conjunction with examples.
[0018] 100 micrograms of chicken ovalbumin (Ovalbumin) and sucrase (Invertase) were subjected to endoglycosidase hydrolysis labeling ( 18 O water buffer solution) and endoglycosidase enzymolysis without labeling (in ordinary ultrapure water buffer solution), and after the enzymolysis sample was purified by ultrafiltration tube and graphitized carbon column in sequence, it was placed in a freeze dryer dry.
[0019] Redissolve the samples with 10-50 microliters of 50% acetonitrile and 0.1% trifluoroacetic acid in water, and take 1-5 microliters for detection by biological mass spectrometry. Both MALDI source and ESI source can be used, and the resolution of mass spectrometry must be greater than 5000 . Obtain unlabeled sugar chain peak intensity (I a ) and its isotopic peak intensity plus 2 Daltons (I a+2 ), and the labeled sugar chain peak intensity (I b ) and its...
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