Preparation and characterization of nickel nanoparticles for binding to his-tag proteins and antigens
Preparation and characterization of nickel nanoparticles for binding to his-tag proteins and antigens
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DOI：
10.1007/s11095-006-9154-7
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发表时间：
2007-02-01
影响因子：
3.7
通讯作者：
Mumper, Russell J.
中科院分区：
文献类型：
--
作者：
Patel, Jigna D.;O'Carra, Ronan;Mumper, Russell J.
目的.这些研究的目的是制备具有少量表面螯合的镍的纳米颗粒（NP），用于获得与非组氨酸标记的蛋白质结合到带电纳米颗粒相比，组氨酸标记的（his-tag）蛋白质的增强的结合。使用乳化蜡、3 mM Brij 78和0. ImM DOGS-NTA-Ni脂质从水包油微乳液前体制备NP（称为Ni-NP）。通过原子发射光谱法（AES）定量纳米颗粒中截留的脂质的量。研究Ni-NP与两种his-标签蛋白（绿色荧光蛋白（GFP）和his-标签HIV-1 Gag p24）的结合。进行小鼠体内研究以评价结合至Ni-NP的His标签Gag p24获得的免疫应答。AES研究表明，约5%的所用DOGS-NTA-Ni脂质被截留在NP中。发现his标签GFP和his标签Gag p24与Ni-NP的最佳结合比率分别为1：33.7和1：35.4w/w。这种相互作用在37 ° C下在pH 7.4的PBS中稳定4小时，并且与表面上没有Ni制备的对照NP（NTA-NP）相比，his-标签GFP与Ni-NP的相互作用增强。体内研究表明，与用明矾佐剂或吸附在对照NTA-NP表面上的蛋白质相比，与Ni-NP结合的His标签Gag p24的血清IgG和IgG 2a应答增强。Ni-NP可用于与His标签蛋白强烈结合。该系统被证明在疫苗递送中具有潜在的应用，用于增强对基于蛋白质的疫苗的免疫应答。
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 degrees 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.