Recent Findings in Peptides and Proteins Described by Researchers from North Carolina State University (The use of a xylosylated plant glycoprotein as an internal standard accounting for N-linked glycan cleavage and sample preparation ...)
By a News Reporter-Staff News Editor at Life Science Weekly -- Investigators discuss new findings in Proteins. According to news reporting originating from Raleigh, North Carolina, by NewsRx correspondents, research stated, "Traditionally, free oligosaccharide internal standards are used to account for variability in glycan relative quantification experiments by mass spectrometry. However, a more suitable internal standard would be a glycoprotein, which could also control for enzymatic cleavage efficiency, allowing for more accurate quantitative experiments."
Our news editors obtained a quote from the research from North Carolina State University, "Hydrophobic, hydrazide N-linked glycan reagents (both native and stable-isotope labeled) are used to derivatize and differentially label N-linked glycan samples for relative quantification, and the samples are analyzed by a reversed-phase liquid chromatography chip system coupled online to a Q-Exactive mass spectrometer. The inclusion of two internal standards, maltoheptaose (previously used) and horseradish peroxidase (HRP) (novel), is studied to demonstrate the effectiveness of using a glycoprotein as an internal standard in glycan relative quantification experiments. HRP is a glycoprotein containing a xylosylated N-linked glycan, which is unique from mammalian N-linked glycans. Thus, the internal standard xylosylated glycan could be detected without interference to the sample. Additionally, it was shown that differences in cleavage efficiency can be detected by monitoring the HRP glycan. In a sample where cleavage efficiency variation is minimal, the HRP glycan performs as well as maltoheptaose. Because the HRP glycan performs as well as maltoheptaose but is also capable of correcting and accounting for cleavage variability, it is a more versatile internal standard and will be used in all subsequent biological studies."
According to the news editors, the research concluded: "Because of the possible lot-to-lot variation of an enzyme, differences in biological matrix, and variable enzyme activity over time, it is a necessity to account for glycan cleavage variability in glycan relative quantification experiments."
For more information on this research see: The use of a xylosylated plant glycoprotein as an internal standard accounting for N-linked glycan cleavage and sample preparation variability. Rapid Communications In Mass Spectrometry, 2013;27(12):1354-8. (Wiley-Blackwell - www.wiley.com/; Rapid Communications In Mass Spectrometry - onlinelibrary.wiley.com/journal/10.1002/(ISSN)1097-0231)
The news editors report that additional information may be obtained by contacting S.H. Walker, WM Keck Fourier Transform Mass Spectrometry Laboratory, Dept. of Chemistry, North Carolina State University, Raleigh, NC 27695, United States. Additional authors for this research include A.D. Taylor and D.C Muddiman (see also Proteins).
Keywords for this news article include: Raleigh, United States, Glycoproteins, North Carolina, Glycoconjugates, North and Central America.
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