On-line preconcentration of proteins in capillary electrochromatography using PLOT-columns based on hyperbranched polymers

  • Elena A. Bessonova the docent of organic chemistry department of Chemical faculty, St. Petersburg state university, St. Petersburg, e-mail: bessonova.Elena.a@gmail.com
  • Vlada U. Koroleva the student of analytic chemistry department of chemical faculty, St. Petersburg state university, St. Petersburg
  • Lyudmila A. Kartsova Dr.Sc.Chem. the professor of organic chemistry department of Chemical faculty, St. Petersburg state university, St. Petersburg
  • Vera E. Potolitsyna the post-graduate student of analytic chemistry department of chemical faculty, St. Petersburg state university, St. Petersburg
Keywords: capillary electrochromatography, PLOT-columns, hyperbranched polymers, proteins, on-line preconcentration.

Abstract

In our study we have investigated the potential of using of new water-soluble oligosaccharide
derivatives hyperbranched polyethylenimin with molecular weight 25 kDa and different degree of
modification maltose (PEI-Mal A - 77% modified, PEI-Mal C - 32% modification), as the stationary phase in
capillary electrochromatography (CEC) for the separation and concentration of proteins (albumin, insulin,
myoglobin, lysozyme). It was synthesized columns with thin porous layer of sorbent on the inner surface of
the fused silica capillary (PLOT-column) based on the hyperbranched polymer. We have investigated the
possibility of different techniques of on-line preconcentration of analytes to reduce the detection limits of
analytes in CEC. We have estimated comparative parameters such as detection limits, efficiency and
separation selectivity. It was shown that the combination of sample stacking with electrokinetic injection
(FESI) and large volume sample sacking allows achieving the concentration factors up to 1000. Established
principles have been tested on real objects (urine, blood serum).

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References

1.Seiler M. Hyperbranched polymers: phase behavior and new applications in the field of
chemical engineering // Fluid Phase Equilibria. 2006. V. 241. P. 155-174.
2.Kolhe P., Misra E., Kannan R.M. et al. Drug complexation, in vitro release and cellular
entry of dendrimers and hyperbranched polymers // International J. of Pharmaceutics.
2003. V. 259. P. 143-160.
3.Klajnert B., Appelhans D. et al. The influence of densely organized maltose shells on
the biological properties of poly(propylene imine) dendrimers: new effects dependent on
hydrogen bonding // Chem. Eur. J. 2008. V. 14. P. 7036-7041.
4.Azagarsamy M. A., Yesilyurt V., Thayumanavan S. Disassembly of dendritic micellar
containers due to protein binding // J. Am. Chem. Soc. 2010. V. 132. P. 4550-4551.
5.Pedziwiatr E., Shcharbin D., Chonco L. et al. Binding properties of water-soluble
carbosilane dendrimers // J. Fluoresc. 2009. V. 19. P. 267-275.
6.Shcharbin D., Janicka M., Wasiak M. et al. Serum albumins have five sites for binding
of cationic dendrimers // Biochim. Biophys. Acta. 2007. V. 1774. P. 946-951.
7.Stathakis C., Arriaga E.A., Dovichi N.J. Cationic and anionic polymeric additives for
wall deactivation and selectivity control in the capillary electrophoretic separation of
proteins in food samples // J. Chromatogr. A. 1998. V. 817. P. 227-232.
8.Liu Q., Tian J., Zhang C., Yang H. et al. Cationic poly(amidoamine) dendrimers as
additives for capillary electroseparation and detection of proteins // Electrophoresis. 2011.
V. 32. P. 1302-1308.
9.Montealegre C., Rasines B., Gomez R. et al. Characterization of carboxylateterminated
carbosilane dendrimers and their evaluation as nanoadditives in capillary
electrophoresis for vegetable protein profiling // J. Chromatogr. A. 2012. V. 1234. P. 16-
21.
10. Boonyakong C., Tucker S.A. Capillary electrophoresis using core-based
hyperbranched polyethyleneimine (CHPEI) static-coated capillaries // J. Sep. Sci. 2009. V.
32. P. 3489-3496.
11. Shou C., Zhang Z. Preparation and characterization of hyperbranched polyester
capillary columns used for the separation of basic proteins // J. of Applied Polymer
Science. 2009. V. 111. P. 2141-2147.
12. Shou C., Song N., Zhang Z. Synthesis of hyperbranched poly(3-methyl-3-
hydroxymethyloxetane) and their application to separate basic proteins by adsorption
coated column // J. of Applied Polymer Science. 2010. V. 116. P. 2473-2479.
13. Appelhans D., Komber H., Abdul Quadir M. et al. Hyperbranched PEI with various
oligosaccharide architectures: Synthesis, characterization, ATP complexation and cellular
uptake // J. Biomacromolecules 2009. V. 10. P. 1114-1124.
14. Burgi D.S., Chien R.L. Improvement in the method of sample stacking for gravity
injection in capillary zone electrophoresis // Analytical Biochemistry. 1992. V. 202.
P. 306-309.
15. Бессонова Е.А., Поликарпов Н.А., Карцова Л.А. и др. Исследование
возможностей новых сверхразветвленных полимеров в качестве
псевдостационарных фаз в электрокинетической хроматографии при определении
белков //Ж. Вестник СПбГУ. Сер. 4. 2011. Вып. 1. С. 100-106.
Published
2019-11-18
How to Cite
Bessonova, E. A., Koroleva, V. U., Kartsova, L. A., & Potolitsyna, V. E. (2019). On-line preconcentration of proteins in capillary electrochromatography using PLOT-columns based on hyperbranched polymers. Sorbtsionnye I Khromatograficheskie Protsessy, 14(2). Retrieved from https://journals.vsu.ru/sorpchrom/article/view/1466