Electrophoresis

George D. J. Phillies

doi:10.1017/CBO9780511843181.004

3 - Electrophoresis

Published online by Cambridge University Press: 05 August 2012

George D. J. Phillies

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George D. J. Phillies: Affiliation:
Worcester Polytechnic Institute, Massachusetts

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Introduction

The early electrophoresis experiments of Tiselius, first published in 1930, examined the motions of proteins in bulk solution as driven by an applied electrical field(1). In the original method, a mixture of proteins began at a fixed location. Under the infiuence of the field, different protein species migrated through solution at different speeds. In time, the separable species moved to distinct locations (“bands”). Electrophoresis is now a primary technique for biological separations(2, 3). Two improvements were critical to establishing the central importance of electrophoresis in biochemistry: First, thin cells and capillary tubes replaced bulk solutions. Second, gels and polymer solutions replaced the simple liquids used by Tiselius as support media. These two improvements greatly increased the resolution of an electrophoretic apparatus. Electrophoresis in true gels is a long-established experimental method. The use of polymer solutions as support media is more recent. An earlymotivation for their use was the suppression of convection, but electrophoretic media that enhance selectivity via physical or chemical interaction with migrating species are now an important biochemical tool.

Electrophoresis and sedimentation have a fundamental similarity: in each method, one observes how particular molecules move when an external force is applied to them. In sedimentation, the enhancement of buoyant forces by the ultracentrifuge causes macromolecules to settle or rise. In electrophoresis, the applied electrical field causes charged macromolecules to migrate. The experimental observable is the drift velocity of the probe as one changes the molecular weight and concentration of the matrix, the size or shape of the probe, or the strength of the external force.

Type: Chapter
Information: Phenomenology of Polymer Solution Dynamics , pp. 30 - 68

DOI: https://doi.org/10.1017/CBO9780511843181.004 [Opens in a new window]

Publisher: Cambridge University Press

Print publication year: 2011

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References

[1] A., Tiselius. The moving-boundary method of studying the electrophoresis of proteins. Nova Acta Regiae Societatis Scientiarum Upsaliensis Ser. IV, 7 (1930), 4.Google Scholar

[2] D. R., Baker. Capillary Electrophoresis, (New York, NY: John Wiley and Sons, 1995).Google Scholar

[3] P. D., Grossman and J. C., Colburn. Capillary Electrophoresis: Theory and Practice, (San Diego, CA: Academic Press, 1992).Google Scholar

[4] J.-L., Viovy. Electrophoresis of DNAand other polyelectrolytes: Physical mechanisms. Revs. Mod. Phys., 72 (2000), 813–872.Google Scholar

[5] A. E., Barron, W. M., Sunada, and H. W., Blanch. Capillary electrophoresis of DNA in uncrosslinked polymer solutions: Evidence for a new mechanism of DNA separation. Biotech. and Bioeng., 52 (1996), 259–270.Google Scholar

[6] A. E., Barron, W. M., Sunada, and H. W., Blanch. The effects of polymer properties on DNA separations by capillary electrophoresis in uncross-linked polymer solutions. Electrophoresis, 17 (1996), 744–757.Google Scholar

[7] S. P., Radko and A., Chrambach. Electrophoretic migration of submicron polystyrene latex spheres in solutions of linear polyacrylamide. Macromolecules, 32 (1999), 2617–2628.Google Scholar

[8] D., Langevin and F., Rondelez. Sedimentation of large colloidal particles through semidilute polymer solutions. Polymer, 19 (1978), 875–882.Google Scholar

[9] A. G., Ogston, B. N., Preston, and J. D., Wells. On the transport of compact particles through solutions of chain-polymers. Proc. Roy. Soc. London (A), 333 (1973), 297–316.Google Scholar

[10] K. A., Streletzky and G. D. J., Phillies. Translational diffusion of small and large mesoscopic probes in hydroxypropylcellulose-water in the solutionlike regime. J. Chem. Phys., 108 (1998), 2975–2988.Google Scholar

[11] K. A., Streletzky and G. D. J., Phillies. Relaxational mode structure for optical probe diffusion in high molecular weight hydroxypropylcellulose. J. Polym. Sci. B, 36 (1998), 3087–3100.Google Scholar

[12] S., Saha, D. M., Heuer, and L. A., Archer. Effect of matrix chain length on the electrophoretic mobility of large linear and branched DNA in polymer solutions. Electrophoresis, 25 (2004), 396–404.Google Scholar

[13] D. M., Heuer, S., Saha, and L. A., Archer. Electrophoretic dynamics of large DNA stars in polymer solutions and gels. Electrophoresis, 24 (2003), 3314–3322.Google Scholar

[14] C., Heller. Finding a universal low-viscosity polymer for DNA separation. Electrophoresis, 19 (1998), 1691–1698.Google Scholar

[15] C., Carlsson, A., Larsson, A., Jonsson, and B., Norden. Dancing DNA in capillary solution electrophoresis. J. Am. Chem. Soc., 117 (1995), 3871–3872.Google Scholar

[16] X., Shi, R. W., Hammond, and M. D., Morris. DNA conformational dynamics in polymer solutions above and below the entanglement limit. Anal. Chem., 67 (1995), 1132–1138.Google Scholar

[17] O., Carmejane, Y., Yamaguchi, T. I., Todorov, and M. D., Morris. Three-dimensional observation of electrophoretic migration of dsDNA in semidilute hydroxyethylcellulose solution. Electrophoresis, 22 (2001), 2433–2441.Google Scholar

[18] W. M., Sunada and H. W., Blanch. Microscopy of DNA in dilute polymer solutions. Biotechnol. Progr., 14 (1998), 766–772.Google Scholar

[19] R. W., Hammond, X., Shi, and M. D., Morris. Dynamics of T2 DNAduring electrophoresis in entangled and ultradilute hydroxyethyl cellulose solutions. J. Microcolumn Separations, 8 (1996), 201–210.Google Scholar

[20] M., Ueda, H., Oana, Y., Baba, M., Doi, and K., Yoshikawa. Electrophoresis of long DNA molecules in linear polyacrylamide solutions. Biophys. Chem., 71 (1998), 113–123.Google Scholar

[21] T. N., Chiesl, R. E., Forster, B. E., Root, M., Larkin, and A. E., Barron. Stochastic single-molecule videomicroscopy methods to measure electrophoretic DNA migration modalities in polymer solutions above and below entanglement. Anal. Chem., 79 (2007), 7740–7747.Google Scholar

[22] N. C., Seeman. Nucleic-acid junctions and lattices. J. Theor. Biol., 99 (1982), 237–247.Google Scholar

[23] The order Teuthida (or Teuthoida or Tenthoidea) are the squids, the largest order of marine cephalopods. See Integrated Taxonomic Information System http://www.itis.gov.

[24] G. D. J., Phillies. Elementary Lectures in Statistical Mechanics, (New York, NY: Springer-Verlag, 2000).Google Scholar

[25] J., Skolnick and A., Kolinski. Dynamics of dense polymer systems. Computer simulations and analytic theories. Adv. Chem. Phys., 78 (1989), 223–278.Google Scholar

[26] L., Mitnik, L., Salome, J. L., Viovy, and C., Heller. Systematic study of field and concentration effects in capillary electrophoresis of DNAin polymer solutions. J. Chromatogr. A, 710 (1995), 309–321.Google Scholar

[27] C., Heller. Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism. Electrophoresis, 20 (1999), 1962–1977.Google Scholar

[28] A. E., Nkodo and B., Tinland. Simultaneous measurements of the electrophoretic mobility, diffusion coefficient and orientation of dsDNA during electrophoresis in polymer solutions. Electrophoresis, 23 (2002), 2755–2765.Google Scholar

[29] M. R., Karim, J.-C., Janson, and T., Takagi. Size-dependent separation of proteins in the presence of sodium dodecyl sulfate and dextran in capillary electrophoresis: Effect of molecular weight of dextran. Electrophoresis, 15 (1994), 1531–1534.Google Scholar

[30] T., Takagi and M. R., Karim. A new mode of size-dependent separation of proteins by capillary electrophoresis in presence of sodium dodecyl sulfate and concentrated oligomeric dextran. Electrophoresis, 16 (1996), 1463–1467.Google Scholar

[31] M., Nakatani, A., Shibukawa, and T., Nakagawa. Separation mechanism of pullulan solution-filled capillary electrophoresis of sodium dodecyl sulfate-proteins. Electrophoresis, 17 (1996), 1584–1586.Google Scholar

[32] D. B., Gomis, S., Junco, Y., Exposito, and M., Gutierrez. Size-based separations of proteins by capillary electrophoresis using linear polyacrylamide as a sieving medium: Model studies and analysis of cider proteins. Electrophoresis, 24 (2003), 1391–1396.Google Scholar

[33] G., Oliver, C., Simpson, M. B., Kerby, A., Tripathi, and A., Chauhan. Electrophoretic migration of proteins in semidilute polymer solutions. Electrophoresis, 29 (2008), 1152–1163.Google Scholar

[34] M., Stastna, S. P., Radko, and A., Chrambach. Capillary zone electrophoresis of proteins in semidilute polymer solutions: Inter- and intra-polymer predictability of size-dependent retardation. Electrophoresis, 20 (1999), 2884–2890.Google Scholar

[35] A., Chrambach and S. P., Radko. Size-dependent retardation and resolution by electrophoresis of rigid, submicron-sized particles, using buffered solutions in the presence of polymers: A review of recent work from the authors' laboratory. Electrophoresis, 21 (2000), 259–265.Google Scholar

[36] S. P., Radko and A., Chrambach. Mechanisms of retardation of rigid spherical particles with 3 to 1,085 nm radius in capillary electrophoresis, using buffered polyacrylamide (molecular weight 5 × 106) solutions. Electrophoresis, 17 (1996), 1094–1102.Google Scholar

[37] S. P., Radko and A., Chrambach. Capillary zone electrophoresis of rigid submicron size particles in polyacrylamide solution. Selectivity, peak spreading and resolution. J. Chromatography A, 848 (1999), 443–455.Google Scholar

[38] D. W., Armstrong, G., Schulte, J. M., Schneiderheinze, and D. J., Westenberg. Separating microbes in the manner of molecules. 1. Capillary electrokinetic approaches. Anal. Chem., 71 (1999), 5465–5469.Google Scholar

[39] A. R., Altenberger and M., Tirrell. On the theory of self-diffusion in a polymer gel. J. Chem. Phys., 80 (1984), 2208–2213.Google Scholar

[40] R. I., Cukier. Diffusion of Brownian spheres in semidilute polymer solutions. Macromolecules, 17 (1984), 252–255.Google Scholar

[41] D., Rodbard and A., Chrambach. Unified theory for gel electrophoresis and gel filtration. Proc. Natl. Acad. Sci. USA, 65 (1970), 970–977.Google Scholar

[42] S. P., Radko and A., Chrambach. Mechanistic insights derived from retardation and peak broadening of particles up to 200 nm in diameter in electrophoresis in semidilute polyacrylamide solutions. Electrophoresis, 19 (1998), 2423–2431.Google Scholar

[43] C., Wu, T., Liu, B., Chu, D. K., Schneider, and V., Graziano. Characterization of the PEO-PPO-PEO triblock copolymer and its application as a separation medium in capillary electrophoresis. Macromolecules, 30 (1997), 4574–4583.Google Scholar

[44] C., Wu, T., Liu, and B., Chu. Viscosity-adjustable block copolymer for DNA separation by capillary electrophoresis. Electrophoresis, 19 (1998), 231–241.Google Scholar

[45] R. L., Rill, Y., Liu, D. H., Van Winkle, and B. R., Locke. Pluronic copolymer liquid crystals: unique, replaceable media for capillary gel electrophoresis. J. Chromatogr. A, 817 (1998), 287–295.Google Scholar

[46] R. L., Rill, B. R., Locke, Y., Liu, and D. H., Van Winkle. Electrophoresis in lyotropic polymer liquid crystals. Proc. Natl. Acad. Sci. USA, 95 (1998), 1534–1539.Google Scholar

[47] R. L., Rill, and M. A., Al-Sayah. Peptide separations by slab gel electrophoresis in pluronic F127 polymer liquid crystals. Electrophoresis, 25 (2004), 1249–1254.Google Scholar

[48] S., Menchen, B., Johnson, M. A., Winnik, and B., Xu. Flowable networks as DNA sequencing media in capillary columns. Electrophoresis, 17 (1996), 1451–1459.Google Scholar

[49] E., Stellwagen, J. D., Prantner, and N. C., Stellwagen. Do zwitterions contribute to the ionic strength of a solution?Analyt. Biochem., 373 (2008), 407–409.Google Scholar

[50] G., Scatchard and J. G., Kirkwood. Das Verhalten von Zwitterionen und von mehrwertigen Ionen mit weit entfernten Ladungen in Elektrolytlosungen. Physikalische Zeit., 33 (1932), 297–300.Google Scholar

[51] J. G., Kirkwood. Theory of solutions of molecules containing widely separated charges with special application to zwitterions. J. Chem. Phys., 2 (1934), 351–361.Google Scholar

[52] K. S., Schmitz. Quasi-elastic scattering by biopolymers. 6. Diffusion of mononucleosomes and oligonucleosomes in the presence of static and sinusoidal electric-fields. Biopolymers, 21 (1982), 1383–1398.Google Scholar

[53] K. S., Schmitz. Quasi-elastic light scattering studies on T7 DNA in the presence of a sinusoidal electric field. Chem. Phys., 79 (1982), 297–305.Google Scholar

[54] T., Imaeda, Y., Kimura, K., Ito, and R., Hayakawa. New formulation for data analysis in the quasielastic light scattering with the sinusoidal electric field and its application to the spherical polyions in aqueous solutions. J. Chem. Phys., 101 (1994), 950–954.Google Scholar

[55] K., Ito, S., Ooi, N., Nishi, Y., Kimura, and R., Hayakawa. New measurement method of the autocorrelation function in the quasielastic light scattering method with the sinusoidal electric field. J. Chem. Phys., 100 (1993), 6098–6100.Google Scholar

[56] K., Ito, S., Ooi, Y., Kinura, and R., Hayakawa. New measurement method for the quasielastic light scattering with the sinusoidal electric field by use of an extended Wiener-Khinchin theorem. J. Chem. Phys., 101 (1994), 4463–4465.Google Scholar

[57] K., Ito and R., Hayakawa. Quasi-elastic light scattering with the sinusoidal electric field: New measurement methods and frequency dispersion of the electrophoretic mobility and diffusion constant of polyions. Colloids and Surfaces A, 148 (1999), 135–148.Google Scholar

[58] G. D. J., Phillies. Bispectral analysis as a probe of quasi-elastic light-scattering intensity fluctuations. J. Chem. Phys., 72 (1980), 6123–6133.Google Scholar

[59] G. D. J., Phillies. Correction: Bispectral analysis as a probe of quasi-elastic light-scattering intensity fluctuations. J. Chem. Phys., 74 (1981), 5333.Google Scholar

[60] T.-H., Lin. Diffusion of TiO2 particles through a poly(ethylene oxide) melt. Makromol. Chem., 187 (1986), 1189–1196.Google Scholar

[61] G. W., Slater, C., Desruisseaux, S. J., Hubert, et al. Theory of DNA electrophoresis: A look at some current challenges. Electrophoresis, 21 (2000), 3873–3887.Google Scholar

[62] T. C., Laurent, I., Bjork, A., Pietruszkiewicz, and H., Persson. On the interaction between polysaccharides and other macromolecules. II. The transport of globular particles through hyaluronic acid solutions. Biochim. Biophys. Acta, 78 (1963), 351–359.Google Scholar

[63] Z., Bu and P. S., Russo. Diffusion of dextran in aqueous hydroxypropylcellulose. Macromolecules, 27 (1994), 1187–1194.Google Scholar

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