Quantitative Characterization of Site-Specific Glycosylation of Glycoproteins – Using Human transferrin as a Model

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	Quantitative Characterization of Site-Specific Glycosylation of Glycoproteins – Using Human transferrin as a Model
	Wei, R.; Ang, I. L. ; Lam, M. M. T.; Zhang, P.; Lei, K. ; Poon, C. W.
	2018-06-08
Source Publication	5th Macau Symposium on Biomedical Sciences 2018
Abstract	Background: Glycoproteins carrying aberrant glycosylations, such as carbohydrate deficient transferrin, have been used as disease biomarkers. Deciphering detailed information of site-specific glycosylation not only can help us understanding the structural and functional relationship of glycoproteins, but also can create new opportunities in disease diagnostics. Objective and Method: Use human transferrin as a glycoprotein model, we aimed to identify experimental factors that affect mass spectrometry-based quantitative characterization of carbohydrate sides at the individual glycosylation sites, and further to develop a new method that can be used to quantify the degree of sialylation and fucosylation of human transferrin in a label-free and antibody-free manner. Results and Discussion: We observed that protease, desalting tips and different digestion protocols can significantly affect the relative signal intensities of individual glycosylated peptides released from transferrin. Systematically evaluate the individual experimental factor can alleviate such quantification bias. Based on the optimized conditions, we invented a new method to quantify the degree of sialylation and fucosylation of human transferrin in a single assay. Conclusion: We have illustrated that without identifying each step of the sample preparation, which may lead to release glycopeptides containing different glycosylation sites in a biased manner. Carefully optimization of the digestion protocol for protein digestion can help alleviating the quantification bias due to digestion bias. Quantification of different glycosylation forms in one single assay may have high potential to provide more information and provide a new sight for human transferrin related disease diagnostics.
Keyword	glycoprotein proteomics glycosylation glycopeptide mass spectrometry
Language	英語English
The Source to Article	PB_Publication
PUB ID	41813
Document Type	Conference paper
Collection	DEPARTMENT OF BIOMEDICAL SCIENCES
Corresponding Author	Poon, C. W.
Recommended Citation GB/T 7714	Wei, R.,Ang, I. L. ,Lam, M. M. T.,et al. Quantitative Characterization of Site-Specific Glycosylation of Glycoproteins – Using Human transferrin as a Model[C], 2018.
APA	Wei, R.., Ang, I. L. ., Lam, M. M. T.., Zhang, P.., Lei, K. ., & Poon, C. W. (2018). Quantitative Characterization of Site-Specific Glycosylation of Glycoproteins – Using Human transferrin as a Model. 5th Macau Symposium on Biomedical Sciences 2018.

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