Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Open Space Laboratory C-2, Tsukuba, Ibaraki, Japan.
Electrophoresis. 2013 Aug;34(16):2432-9. doi: 10.1002/elps.201300154. Epub 2013 Jul 26.
Supported molecular matrix electrophoresis (SMME), in which a hydrophilic polymer such as PVA serves as a support within a porous PVDF membrane, was recently developed. This method is similar to cellulose acetate membrane electrophoresis but differs in the compatibility to glycan analysis of the separated bands. In this report, we describe the first instance of the application of SMME to human serum fractionation, and demonstrate the differences with serum fractionation by cellulose acetate membrane electrophoresis. The SMME membrane exhibited almost no EOF during electrophoresis, unlike the cellulose acetate membrane, but afforded comparative results for serum fractionation. The visualization of each fraction was achieved by conventional staining with dye such as Direct Blue-71, and objective quantification was obtained by densitometry after inducing membrane transparency with 1-nonene. Immunostaining was also achieved. Moreover, mass spectrometric analysis of both N-linked and O-linked glycans from the separated bands was demonstrated. Serum fractionation and glycan profiling of each fraction using SMME will enable novel insights into the relationships between various glycosylation profiles and disease states.
最近开发了一种支持的分子基质电泳（SMME），其中亲水性聚合物如 PVA 作为多孔 PVDF 膜内的支撑物。这种方法类似于醋酸纤维素膜电泳，但在分离带的聚糖分析的兼容性上有所不同。在本报告中，我们描述了 SMME 首次应用于人血清分离，并展示了与醋酸纤维素膜电泳的血清分离的差异。与醋酸纤维素膜不同，SMME 膜在电泳过程中几乎没有 EOF，但对血清分离提供了相当的结果。通过用直接蓝 71 等染料对每个馏分进行常规染色，可以实现可视化，并且通过用 1-壬烯诱导膜透明度后进行密度计法进行客观定量。还实现了免疫染色。此外，还展示了从分离带中 N-连接和 O-连接聚糖的质谱分析。使用 SMME 进行血清分离和各馏分的聚糖分析将使我们能够深入了解各种糖基化谱与疾病状态之间的关系。
