title: Conic electrophoretic concentrator for charged macromolecules.
abstract: 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 12-15 min by a factor of ∼100,000 with the total yield of 60-80%. 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's surface can be further revealed by specific reaction with a microarray of antibody molecules. Such "reversed magnetic array" 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.
authors.affiliation: Institute of Theoretical and Experimental Biophysics, Russian Academy of Science, Pushchino, Moscow Region, Russia. vmorozov@gmu.edu
authors.forename: Victor N
authors.initials: VN
authors.lastname: Morozov
authors.forename: Yuri M
authors.initials: YM
authors.lastname: Shlyapnikov
authors.forename: Jessica
authors.initials: J
authors.lastname: Kidd
authors.forename: Tamara Y
authors.initials: TY
authors.lastname: Morozova
authors.forename: Elena A
authors.initials: EA
authors.lastname: Shlyapnikova
mesh_terms: D000818:Animals; D001944:Breath Tests; D003956:Dialysis / Q000191:economics / Q000295:instrumentation; D004586:Electrophoresis / Q000191:economics / Q000295:instrumentation*; D004867:Equipment Design; D006801:Humans; D059346:Magnets / Q000737:chemistry; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results
chemical_list: D011506:Proteins