Conic Electrophoretic Concentrator for Charged Macromolecules

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• 作者：Victor N. Morozov ; Yuri M. Shlyapnikov ; Jessica Kidd ; Tamara Y. Morozova ; Elena A. Shlyapnikova
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：July 15, 2011
• 年：2011
• 卷：83
• 期：14
• 页码：5548-5555
• 全文大小：914K
• 年卷期：v.83,no.14(July 15, 2011)
• ISSN：1520-6882

文摘

A simple, rapid, and highly effective technique for concentrating charged macromolecules is described which employs electrophoresis in a conic cell made of a dialysis membrane. The cell is partly submerged in electrolyte solution, and the level of solution slowly moves down during the process. The electric field within the cell is at its maximum in the area that is level with the surface of the external solution. This maximum value increases and its location moves downward following the decreasing level of external solution carrying downward and concentrating charged macromolecules. It has been demonstrated that proteins can be concentrated within 12鈥?5 min by a factor of 100鈥?00 with the total yield of 60鈥?0%. Concentrated proteins can be harvested from the nanoliter-sized cul-de-sac of the conic concentrator using chemically activated magnetic beads. The presence of certain protein molecules linked to the bead鈥檚 surface can be further revealed by specific reaction with a microarray of antibody molecules. Such 鈥渞eversed magnetic array鈥?format was applied to a cone-concentrated exhaled breath condensate (EBC) to reveal the presence of human immunoglobulin in the EBC and to estimate its concentration. The technique may be used for concentrating and detecting trace amounts of pathogens and toxins, in protein crystallization, and in many other applications.