Abstract
Capillary electrophoresis (CE) is one of the most powerful techniques for the separation of biomolecules. However, the separation efficiency of proteins in CE is often compromised by their tendency to interact with the silanol groups on the surface of the inner capillary and by an uncontrolled electroosmotic flow. Herein, we report on the use of novel hydrophilic polymeric coatings that can modulate the properties of the capillary walls. The novelty of these poly(N,N-dimethylacrylamide)-based copolymers relies on the simultaneous presence of chemically reactive groups (N-acryloyloxysuccinimide and glycidyl methacrylate) and silane groups in the backbone, which results in highly stable films due to the covalent reaction between the polymer and the glass silanols. A careful optimization of monomer concentration confers anti-fouling properties to the polymer coatings, and thus allows for highly efficient acidic and alkaline protein separations. Furthermore, the presence of these monomers makes it possible to modulate the electroosmotic flow from negligible to reduced values, depending on the desired application.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Lookhart GL, Bean SR. (2004) Capillary electrophoresis of cereal proteins: an overview. J Capill Electrophor Microchip Technol 9:23–30
Lucy CA, MacDonald AM, Gulcev MD (2008) Non-covalent capillary coatings for protein separations in capillary electrophoresis. J Chromatogr A 1184:81–105
Lauer HH, McManigill D (1986) Capillary zone electrophoresis of proteins in untreated fused silica tubing. Anal Chem 58:166–170
McCormick RM (1988) Capillary zone electrophoretic separation of peptides and proteins using low pH buffers in modified silica capillaries. Anal Chem 60:2322–2328
Green JS, Jorgenson JW (1989) Minimizing adsorption of proteins on fused silica in capillary zone electrophoresis by the addition of alkali metal salts to the buffers. J Chromatogr A 478:63–70
Bachmann S, Vallant R, Bakry R et al (2010) CE coupled to MALDI with novel covalently coated capillaries. Electrophoresis 31:618–629
Zhang Z, Yan B, Liu K et al (2009) CE-MS analysis of heroin and its basic impurities using a charged polymer-protected gold nanoparticle-coated capillary. Electrophoresis 30:379–387
Ullsten S, Zuberovic A, Wetterhall M et al (2004) A polyamine coating for enhanced capillary electrophoresis-electrospray ionization-mass spectrometry of proteins and peptides. Electrophoresis 25:2090–2099
Kašička V (2010) Recent advances in CE and CEC of peptides (2007-2009). Electrophoresis 31:122–146
Liu CY (2001) Stationary phases for capillary electrophoresis and capillary electrochromatography. Electrophoresis 22:612–628
Chiari M, Cretich M, Damin F et al (2000) New adsorbed coatings for capillary electrophoresis. Electrophoresis 21:909–916
Cretich M, Chiari M, Pirri G et al (2005) Electroosmotic flow suppression in capillary electrophoresis: chemisorption of trimethoxy silane-modified polydimethylacrylamide. Electrophoresis 26:1913–1919
Pirri G, Damin F, Chiari M et al (2004) Characterization of a polymeric adsorbed coating for DNA microarray glass slides. Anal Chem 76:1352–1358
Yalçin A, Damin F, Ozkumur E et al (2009) Direct observation of conformation of a polymeric coating with implications in microarray applications. Anal Chem 81:625–630
Cretich M, Reddington A, Monroe M et al (2011) Silicon biochips for dual label-free and fluorescence detection: application to protein microarray development. Biosens Bioelectron 26:3938–3943
Cretich M, Sedini V, Damin F et al (2008) Functionalization of poly(dimethylsiloxane) by chemisorption of copolymers: DNA microarrays for pathogen detection. Sens Actuators B 132:258–264
Cretich M, Sedini V, Damin F et al (2010) Coating of nitrocellulose for colorimetric DNA microarrays. Anal Biochem 397:84–88
Chiari M (2011) Patent application WO 2011124715 A1, silane copolymers and uses thereof, WO 2011124715 A1, 2011.
Sola L, Chiari M (2012) Modulation of electroosmotic flow in capillary electrophoresis using functional polymer coatings. J Chromatogr A 1270:324–329
Marangoni S, Rech I, Ghioni M et al (2010) A 6×8 photon-counting array detector system for fast and sensitive analysis of protein microarrays. Sens Actuators B 149:420–426
Luzinov I, Julthongpiput D, Malz H et al (2000) Polystyrene layers grafted to epoxy-modified silicon surfaces. Macromolecules 33:1043–1048
Williams BA, Vigh G (1996) Fast, accurate mobility determination method for capillary electrophoresis. Anal Chem 68:1174–1180
Mammen M, Dahmann G, Whitesides GM (1995) Effective inhibitors of hemagglutination by influenza virus synthesized from polymers having active ester groups. Insight into mechanism of inhibition. J Med Chem 38:4179–4190
Acknowledgement
This work was financially supported by FP7 EU NADINE Project (contract number 246513) and by Italian Ministry for Research and Education FIRB NEMATIC Project (contract number RBFR 12OOG).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Sola, L., Cretich, M., Chiari, M. (2016). Poly(N,N-Dimethylacrylamide)-Based Coatings to Modulate Electroosmotic Flow and Capillary Surface Properties for Protein Analysis. In: Tran, N., Taverna, M. (eds) Capillary Electrophoresis of Proteins and Peptides. Methods in Molecular Biology, vol 1466. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4014-1_9
Download citation
DOI: https://doi.org/10.1007/978-1-4939-4014-1_9
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-4012-7
Online ISBN: 978-1-4939-4014-1
eBook Packages: Springer Protocols