A nanoparticle-based immobilization assay for prion-kinetics study.
A nanoparticle-based immobilization assay for prion-kinetics study.
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DOI：
10.1186/1477-3155-4-8
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发表时间：
2006-08-17
影响因子：
10.2
通讯作者：
Irudayaraj, Joseph
中科院分区：
文献类型：
--
作者：
Kouassi, Gilles K;Irudayaraj, Joseph
采用亚铁和三氯化铁共沉淀法和微胶束法分别制备了磁性纳米粒子和包金磁性纳米粒子。合成的纳米颗粒在n -羟基琥珀酰亚胺存在下经碳二亚胺活化后被功能化为羧基和氨基酸部分，并作为朊病毒蛋白载体。利用红外光谱（FTIR）和透射电镜（tem）对纳米颗粒的大小和结构进行了表征，证实了重组人朊病毒蛋白（huPrPrec）与纳米颗粒表面的结合。结果表明，随着反应介质中朊病毒浓度的增加，朊病毒的结合率仅略有增加。当蛋白浓度分别为1、2、1.5、2.25和3.57 μ g/ml时，Fe3O4@Au和Fe3O4-LAA的结合速率常数非常相似。当huPrPrec浓度为5 μ g/ml时，Fe3O4-LAA颗粒的结合速率常数更高。该研究为朊蛋白复合物在三维结构上的形成铺平了道路，可以揭示模糊的生理和病理结构以及朊蛋白动力学。
Magnetic and gold coated magnetic nanoparticles were synthesized by co-precipitation of ferrous and ferric chlorides, and by the micromicelles method, respectively. Synthesized nanoparticles were functionalized to bear carboxyl and amino acid moieties and used as prion protein carriers after carbodiimide activation in the presence of N-hydroxysuccinimide. The binding of human recombinant prion protein (huPrPrec) to the surface of these nanoparticles was confirmed by FTIR and the size and structures of the particles were characterized by transmission electron microscopy. Findings indicate that the rate of prion binding increased only slightly when the concentration of prion in the reaction medium was increased. Rate constants of binding were very similar on Fe3O4@Au and Fe3O4-LAA when the concentrations of protein were 1, 2, 1.5, 2.25 and 3.57 microg/ml. For a 5 microg/ml concentration of huPrPrec the binding rate constant was higher for the Fe3O4-LAA particles. This study paves the way towards the formation of prion protein complexes onto a 3-dimensional structure that could reveal obscure physiological and pathological structure and prion protein kinetics.