Czech Academy of Sciences, Institute of Analytical Chemistry, Brno 602 00, Czech Republic.
Czech Academy of Sciences, Institute of Analytical Chemistry, Brno 602 00, Czech Republic.
Anal Chim Acta. 2021 Apr 15;1154:338246. doi: 10.1016/j.aca.2021.338246. Epub 2021 Feb 10.
Polyacrylamide or agarose gels are the most frequently used sieving and stabilizing media in slab gel electrophoresis. Recently, we have introduced a new electrophoretic technique for concentration/separation of milliliter sample volumes. In this technique, the gel is used primarily as an anticonvection media eliminating liquid flow during the electromigration. While serving well for the liquid stabilization, the gels can undergo deformation when exposed to a discontinuous electrolyte buffer system used in epitachophoresis. In this work, we have explored 3D printing to form rigid stabilizing manifolds to minimize liquid flow during the epitachophoresis run. The whole device was printed using the stereolithography technique from a low water-absorbing resin. The stabilizing manifold, serving as the gel substitute, was printed as a replaceable composite structure preventing electrolyte mixing during the separation. Different geometries of the 3D printed stabilizing manifolds were tested for use in concentrating ionic sample components without spatial separation. The presented device can focus analytes from 3 or 4 mL of the sample to 150 μL or less, depending on the collection cup size. With the 150 μL collection cup, this represents the enrichment factor from 20 to 27. The time of concentration was from 15 to 25 min, depending on stabilization media and power used.
聚丙烯酰胺或琼脂糖凝胶是板状凝胶电泳中最常用的筛分和稳定介质。最近,我们引入了一种新的电泳技术,用于浓缩/分离毫升级样品体积。在该技术中,凝胶主要用作抗对流介质,在电泳迁移过程中消除液体流动。虽然凝胶在用于等速电泳的不连续电解质缓冲系统中能很好地稳定液体,但当暴露于该系统时,凝胶会发生变形。在这项工作中,我们探索了 3D 打印来形成刚性稳定流道,以最小化等速电泳过程中的液体流动。整个装置采用立体光刻技术,由低吸水性树脂打印而成。稳定流道作为凝胶替代品,以可更换的复合结构打印,防止分离过程中电解质混合。为了在不进行空间分离的情况下浓缩离子样品成分,测试了不同几何形状的 3D 打印稳定流道。所提出的装置可将来自 3 或 4 mL 样品的分析物浓缩至 150 μL 或更少,具体取决于收集杯的大小。使用 150 μL 收集杯,浓缩因子为 20 到 27 倍。浓缩时间取决于稳定介质和所使用的功率,为 15 到 25 分钟。