Analysis of N-Glycans Profiling from HeLa Cells by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

doi:10.3969/j.issn.0251-0790.2012.06.019

Abstract

Abstract: The structure of N-glycans on cell surface was abnormal in the process of cancer, which was closely related to the invasion and metastasis of tumors. Total glycopeptides were obtained from trace HeLa cells(10⁷) after cell lyses, reduction and acetamidation, trypsin digestion and Oasis-HLB cartridge purification. N-glycans were released by PNGase F digestion and purified by a Sep-Pak C₁₈ cartridge. The structural of N-glycans profiling was first analyzed by positive-ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) after permethylation. The 34 types of N-glycans obtained from HeLa cell included both high-mannose and complex(bi-, tri-, tetra- and penta-antennary) types, some of the latter containing bisecting and Lewis determinants, which were relevant to tumor metastasis in same way. The MALDI-TOF MS technology was suitable for rapid analysis of N-glycans profiling on cancer cell surface in trace amount, in order to find new glycans-makers and more foundation for prevention and diagnosis in early stage of tumors.

Key words: Tumor, HeLa cell, N-glycans, MALDI-TOF MS

CLC Number:

O629.12
O657

TrendMD:

WANG Pei-Pei, YU Guang-Li, JIAO Guang-Ling, ZHAO Xia, LI Jing, CHAI Wen-Gang. Analysis of N-Glycans Profiling from HeLa Cells by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry[J]. Chem. J. Chinese Universities, 2012, 33(06): 1233.

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