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Preparation and Characterization of Nickel Nanoparticles for Binding to His-tag Proteins and Antigens

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Abstract

Purpose

The purpose of these studies was to prepare nanoparticles (NPs) with a small amount of surface-chelated nickel for obtaining enhanced binding of histidine-tagged (his-tag) proteins compared to non-histidine-tagged protein binding to charged nanoparticles.

Materials and Methods

NPs were prepared from oil-in-water microemulsion precursors using emulsifying wax, 3 mM Brij 78 and 0.1 mM DOGS–NTA–Ni lipid (referred to as Ni-NPs). The amount of lipid entrapped in the NPs was quantitated by atomic emission spectroscopy (AES). The Ni-NPs were investigated for binding to two his-tag proteins, green fluorescent protein (GFP) and his-tag HIV-1 Gag p24. In vivo studies in mice were carried out to evaluate the immune responses obtained to his-tag Gag p24 bound to Ni-NPs.

Results

AES studies demonstrated that approximately 5% of the DOGS–NTA–Ni lipid used was entrapped in the NPs. The optimal binding ratio his-tag GFP and his-tag Gag p24 to Ni-NPs was found to be 1:33.7 and 1:35.4 w/w, respectively. This interaction was stable at 37°C in PBS, pH 7.4 over 4 h and the interaction of his-tag GFP with the Ni-NPs was enhanced compared to control NPs prepared with no Ni on the surface (NTA-NPs). The in vivo studies demonstrated enhanced serum IgG and IgG2a responses to his-tag Gag p24 bound to Ni-NPs compared to protein adjuvanted with Alum or adsorbed on the surface of control NTA-NPs.

Conclusions

Ni-NPs can be used to bind strongly to his-tag proteins. This system was demonstrated to have potential applications in vaccine delivery for enhancing immune responses to protein-based vaccines.

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Acknowledgments

This research was funded by NIH-NIAID AI058842 to RJM and JGW. J. Patel was supported, in part, by a Pre-doctoral fellowship received from the American Foundation for Pharmaceutical Education and the 2005 Dissertation Year Fellowship received from the University of Kentucky Graduate School. The authors would like to thank Tricia Coakley in the Environmental Research and Training Laboratory (ERTL) at the University of Kentucky for her technical assistance in analyzing nanoparticle samples by AES.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, 40536-0082, USA

    Jigna D. Patel, Ronan O’Carra & Russell J. Mumper

  2. Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky, Lexington, KY, 40536-0298, USA

    Julia Jones & Jerold G. Woodward

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  1. Jigna D. Patel
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  2. Ronan O’Carra
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Correspondence to Russell J. Mumper.

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Patel, J.D., O’Carra, R., Jones, J. et al. Preparation and Characterization of Nickel Nanoparticles for Binding to His-tag Proteins and Antigens. Pharm Res 24, 343–352 (2007). https://doi.org/10.1007/s11095-006-9154-7

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  • DOI: https://doi.org/10.1007/s11095-006-9154-7

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