Journal of Glycobiology
Open Access

Quantification of O-linked and N-linked glycome in human fibroblast

2^nd Glycobiology World Congress

August 29-31, 2016 Atlanta, USA

Xueli Li

University of Pennsylvania, USA

Scientific Tracks Abstracts: J Glycobiol

Abstract :

Protein glycosylation is increasingly recognized as a crucial modulator of protein function, offering a third layer of biological information over genomics and proteomics. Modern tools for analyzing released N-glycans from cells, the glycome, have shown abnormal protein glycosylation in numerous human diseases. We developed a quantification of glycome in cells. Upon reaching 100% confluence, the cells were washed twice with PBS and harvested using a cell scraper. The cells were then pelleted and washed with PBS by centrifugation. Fibroblast pellets were lysed in more than 200 �¼l PBS, and 200 �¼g protein from the cell lysate was denature and precipitated with 2�� volume of 100% propanol. N-linked and O-linked glycans were released from denatured protein, desalted and permethylated before subject to MALDI-TOF analysis. The quantification of O-linked glycans was achieved by spiking glycans from cell with 25 �¼M of C13-labelled T antigen (m/z 543) and C13-labelled sialylated T antigen (m/z 909). N-linked glycans were quantified using 25 �¼M of C13-labelled Man7GlcNAc2 (m/z 1107). Using this method, we identified abnormal fibroblast glycomes in a number of known patients with congenital disorders of glycosylation and demonstrate cellular glycome as an important tool for diagnosis of these diseases.

Biography :

Xueli Li completed her PhD from Leipzig University, Germany. She is a Research Associate in The Michael J Palmieri Metabolic Laboratory, Children’s Hospital of Philadelphia, Philadelphia, PA. She has more than 8 publications in the area of Glycomics.

Email: LiX3@email.chop.edu