Abstract
Various polysaccharides can successfully migrate through entangled polymer solutions during high-voltage capillary electrophoresis. For neutral polysaccharides, complexation with borate provides the electric charge needed for electromigration, while a fluorescent tag is needed to detect the solute bands with adequate sensitivity. At constant potentials between 50 and 300 V/cm, the charged polysaccharides undergo molecular stretching, resisting the desired separation according to their molecular mass. This problem can be overcome through the use of variable fields, pulsed along the separation capillary at a 180 degree angle. Variables of the pulsing experiment appear to have a profound influence on molecular shape rearrangements of polysaccharides with respect to the separation medium, as demonstrated here with highly efficient separations of polydextrans (8,000-2,000,000 Da).
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Selected References
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- Brassard E., Turmel C., Noolandi J. Observation of orientation and relaxation of protein-sodium dodecyl sulfate complexes during pulsed intermittent field polyacrylamide gel electrophoresis. Electrophoresis. 1991 May;12(5):373–375. doi: 10.1002/elps.1150120509. [DOI] [PubMed] [Google Scholar]
- Carle G. F., Frank M., Olson M. V. Electrophoretic separations of large DNA molecules by periodic inversion of the electric field. Science. 1986 Apr 4;232(4746):65–68. doi: 10.1126/science.3952500. [DOI] [PubMed] [Google Scholar]
- Chen D. Y., Swerdlow H. P., Harke H. R., Zhang J. Z., Dovichi N. J. Low-cost, high-sensitivity laser-induced fluorescence detection for DNA sequencing by capillary gel electrophoresis. J Chromatogr. 1991 Oct 18;559(1-2):237–246. doi: 10.1016/0021-9673(91)80074-q. [DOI] [PubMed] [Google Scholar]
- Cobb K. A., Dolnik V., Novotny M. Electrophoretic separations of proteins in capillaries with hydrolytically stable surface structures. Anal Chem. 1990 Nov 15;62(22):2478–2483. doi: 10.1021/ac00221a013. [DOI] [PubMed] [Google Scholar]
- Cohen A. S., Najarian D. R., Karger B. L. Separation and analysis of DNA sequence reaction products by capillary gel electrophoresis. J Chromatogr. 1990 Sep 7;516(1):49–60. doi: 10.1016/s0021-9673(01)90203-1. [DOI] [PubMed] [Google Scholar]
- Cohen A. S., Najarian D. R., Paulus A., Guttman A., Smith J. A., Karger B. L. Rapid separation and purification of oligonucleotides by high-performance capillary gel electrophoresis. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9660–9663. doi: 10.1073/pnas.85.24.9660. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cumming D. A. Glycosylation of recombinant protein therapeutics: control and functional implications. Glycobiology. 1991 Mar;1(2):115–130. doi: 10.1093/glycob/1.2.115. [DOI] [PubMed] [Google Scholar]
- Demana T., Lanan M., Morris M. D. Improved separation of nucleic acids with analyte velocity modulation capillary electrophoresis. Anal Chem. 1991 Dec 1;63(23):2795–2797. doi: 10.1021/ac00023a023. [DOI] [PubMed] [Google Scholar]
- Ganzler K., Greve K. S., Cohen A. S., Karger B. L., Guttman A., Cooke N. C. High-performance capillary electrophoresis of SDS-protein complexes using UV-transparent polymer networks. Anal Chem. 1992 Nov 15;64(22):2665–2671. doi: 10.1021/ac00046a003. [DOI] [PubMed] [Google Scholar]
- Gassmann E., Kuo J. E., Zare R. N. Electrokinetic separation of chiral compounds. Science. 1985 Nov 15;230(4727):813–814. doi: 10.1126/science.230.4727.813. [DOI] [PubMed] [Google Scholar]
- Grossman P. D., Soane D. S. Experimental and theoretical studies of DNA separations by capillary electrophoresis in entangled polymer solutions. Biopolymers. 1991 Sep;31(10):1221–1228. doi: 10.1002/bip.360311010. [DOI] [PubMed] [Google Scholar]
- Heiger D. N., Cohen A. S., Karger B. L. Separation of DNA restriction fragments by high performance capillary electrophoresis with low and zero crosslinked polyacrylamide using continuous and pulsed electric fields. J Chromatogr. 1990 Sep 7;516(1):33–48. doi: 10.1016/s0021-9673(01)90202-x. [DOI] [PubMed] [Google Scholar]
- Honda S., Makino A., Suzuki S., Kakehi K. Analysis of the oligosaccharides in ovalbumin by high-performance capillary electrophoresis. Anal Biochem. 1990 Dec;191(2):228–234. doi: 10.1016/0003-2697(90)90212-r. [DOI] [PubMed] [Google Scholar]
- Lee K. B., Desai U. R., Palcic M. M., Hindsgaul O., Linhardt R. J. An electrophoresis-based assay for glycosyltransferase activity. Anal Biochem. 1992 Aug 15;205(1):108–114. doi: 10.1016/0003-2697(92)90586-v. [DOI] [PubMed] [Google Scholar]
- Liu J. P., Hsieh Y. Z., Wiesler D., Novotny M. Design of 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde as a reagent for ultrasensitive determination of primary amines by capillary electrophoresis using laser fluorescence detection. Anal Chem. 1991 Mar 1;63(5):408–412. doi: 10.1021/ac00005a004. [DOI] [PubMed] [Google Scholar]
- Liu J. P., Shirota O., Novotny M. Separation of fluorescent oligosaccharide derivatives by microcolumn techniques based on electrophoresis and liquid chromatography. J Chromatogr. 1991 Oct 18;559(1-2):223–235. doi: 10.1016/0021-9673(91)80073-p. [DOI] [PubMed] [Google Scholar]
- Liu J. P., Shirota O., Wiesler D., Novotny M. Ultrasensitive fluorometric detection of carbohydrates as derivatives in mixtures separated by capillary electrophoresis. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2302–2306. doi: 10.1073/pnas.88.6.2302. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu J., Shirota O., Novotny M. V. Sensitive, laser-assisted determination of complex oligosaccharide mixtures separated by capillary gel electrophoresis at high resolution. Anal Chem. 1992 Apr 15;64(8):973–975. doi: 10.1021/ac00032a026. [DOI] [PubMed] [Google Scholar]
- Lumpkin O. J., Déjardin P., Zimm B. H. Theory of gel electrophoresis of DNA. Biopolymers. 1985 Aug;24(8):1573–1593. doi: 10.1002/bip.360240812. [DOI] [PubMed] [Google Scholar]
- Nashabeh W., el Rassi Z. Capillary zone electrophoresis of alpha 1-acid glycoprotein fragments from trypsin and endoglycosidase digestions. J Chromatogr. 1991 Jan 4;536(1-2):31–42. doi: 10.1016/s0021-9673(01)89234-7. [DOI] [PubMed] [Google Scholar]
- Novotny M. V., Cobb K. A., Liu J. P. Recent advances in capillary electrophoresis of proteins, peptides and amino acids. Electrophoresis. 1990 Sep;11(9):735–749. doi: 10.1002/elps.1150110911. [DOI] [PubMed] [Google Scholar]
- Novotny M. V., Sudor J. High-performance capillary electrophoresis of glycoconjugates. Electrophoresis. 1993 May-Jun;14(5-6):373–389. doi: 10.1002/elps.1150140163. [DOI] [PubMed] [Google Scholar]
- Paulson J. C. Glycoproteins: what are the sugar chains for? Trends Biochem Sci. 1989 Jul;14(7):272–276. doi: 10.1016/0968-0004(89)90062-5. [DOI] [PubMed] [Google Scholar]
- Rutishauser U., Acheson A., Hall A. K., Mann D. M., Sunshine J. The neural cell adhesion molecule (NCAM) as a regulator of cell-cell interactions. Science. 1988 Apr 1;240(4848):53–57. doi: 10.1126/science.3281256. [DOI] [PubMed] [Google Scholar]
- Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
- Slater G. W., Rousseau J., Noolandi J., Turmel C., Lalande M. Quantitative analysis of the three regimes of DNA electrophoresis in agarose gels. Biopolymers. 1988 Mar;27(3):509–524. doi: 10.1002/bip.360270311. [DOI] [PubMed] [Google Scholar]
- Soini H., Riekkola M. L., Novotny M. V. Chiral separations of basic drugs and quantitation of bupivacaine enantiomers in serum by capillary electrophoresis with modified cyclodextrin buffers. J Chromatogr. 1992 Sep 11;608(1-2):265–274. doi: 10.1016/0021-9673(92)87133-s. [DOI] [PubMed] [Google Scholar]
- Torii M., Sakakibara K., Alberto B. P., Misaki A. Immunochemical studies on dextrans: microheterogeneity of dextran NRRL B1397 demonstrated by precipitation with concanavalin A. Biochem Biophys Res Commun. 1976 Sep 7;72(1):236–242. doi: 10.1016/0006-291x(76)90985-2. [DOI] [PubMed] [Google Scholar]
- Tsuji K. High-performance capillary electrophoresis of proteins. Sodium dodecyl sulphate-polyacrylamide gel-filled capillary column for the determination of recombinant biotechnology-derived proteins. J Chromatogr. 1991 Jul 26;550(1-2):823–830. doi: 10.1016/s0021-9673(01)88585-x. [DOI] [PubMed] [Google Scholar]
- Viovy J. L. Reptation-breathing theory of pulsed electrophoresis: dynamic regimes, antiresonance and symmetry breakdown effects. Electrophoresis. 1989 May-Jun;10(5-6):429–441. doi: 10.1002/elps.1150100521. [DOI] [PubMed] [Google Scholar]