Abstract
Tumor-associated structural alterations of O-linked glycans in mucins have often been reported. However, mucin characterization has been lagging following adoption of modern techniques such as proteomics because of their large size, polymeric nature, and heterogeneous glycosylation. For mucins to be used as biomarkers, a convenient and high-throughput technology for characterization of mucins including analysis of glycan moieties must first be developed. Supported Molecular Matrix Electrophoresis (SMME) is a membrane electrophoresis in which hydrophilic polymer soaking into a porous membrane such as polyvinylidene difluoride (PVDF) is used as the separation medium. The electrophoretic conditions for cellulose acetate membrane electrophoresis can be applied to SMME without significant modifications. Treatment of the SMME membrane under alkaline β-elimination conditions does not induce degradation products of hexose oligomers that would otherwise interfere with glycan analysis. This technique was applied to the characterization of MUC1 produced by three cancer cell lines (T47D, HPAF-II, and BxPC3).
