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Development and characterization of stabilized, polymerized phospholipid bilayers on silica particles for specific immobilization of His-tagged proteins

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Abstract

Stabilized phospholipid bilayer (PLB) coated silica microspheres were prepared via polymerization of lipid monomers. These lipid coated silica microspheres are stable to both extended storage in solution at 4∘C and dry storage at room temperature. These stabilized lipid coated microspheres have also been functionalized with nickel-chelating lipids, a commonly used tool for immobilizing polyhistidine-tagged proteins. It is shown that 6xHis-EGFP interacts with (poly)bis-SorbPC/DOGS-NTA-Ni2+ coated silica and this interaction was interrupted by washing with imidazole indicating the reversibility of the interaction. No interaction was observed between the functionalized silica substrate and EGFP, which lacks the 6xHis-tag. Furthermore, these biocompatible (poly)bis-SorbPC coated microspheres were able to minimize non-specific protein adsorption.

Biocompatible and highly stable phospholipid bilayers supported on silica microspheres were prepared by UV polymerization of a synthetic lipid. The resulting polymeric coatings were stable to extended storage in both solution and dry conditions. Furthermore, these biocompatible polymeric phospholipid bilayers allow incorporation of biologically and chemically important chemical moieties such as NTA-Ni2+ and exhibited enhanced capability of reducing nonspecific protein adsorption. X-axis in the figure is fluorescence intensity of GFP in a.u.

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Acknowledgements

The author would like to thank the Cellular Imaging Facility Core of the University of Arizona for the flow cytometry experiments.

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  1. Department of Chemistry, Washburn University, Topeka, Kansas, 66621, USA

    SEID M ADEM

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Supplementary Information

Fluorescence images showing: the stability of (poly) bis-SorbPC coated silica microspheres to extended storage at 4∘C and dry storage and acetonitrile washing; functionalization with DOGS-NTA-Ni2+ and non-specific protein adsorption of (poly)bis-SorbPC coated silica microspheres are presented in the supplemental information available at www.ias.ac.in/chemsci

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ADEM, S.M. Development and characterization of stabilized, polymerized phospholipid bilayers on silica particles for specific immobilization of His-tagged proteins. J Chem Sci 127, 729–735 (2015). https://doi.org/10.1007/s12039-015-0829-7

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  • DOI: https://doi.org/10.1007/s12039-015-0829-7

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