Fabrication of Anionic Sulfate-Functionalized Nanoparticles as an Immunosensor by Protein Immobilization
Fabrication of Anionic Sulfate-Functionalized Nanoparticles as an Immunosensor by Protein Immobilization
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DOI：
10.1021/la9043717
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发表时间：
2010-05-18
期刊：
影响因子：
3.9
通讯作者：
Kim, Wan-Joong
中科院分区：
文献类型：
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作者：
Kim, Sanghee;Pyo, Hyeon-Bong;Kim, Wan-Joong
以过硫酸钾(K PS)在四硼酸钠存在下通过乳液聚合热分解制备阴离子硫酸盐(SO(4)(-))功能化聚苯乙烯(PS)纳米颗粒。在 X 射线光电子能谱 (X PS) 分析中，Zeta 电位值为 -40.6 mV，并且 S 2p 光谱向 162.7 eV (2p(3/2)) 和 164.4 eV (2p(1/2)) 附近的较低结合能区域移动，证实了颗粒表面固/液界面上存在 SO.: 基团。人免疫球蛋白 G (hIgG) 和心肌肌钙蛋白 I (c-In I) 的带正电荷的抗体与带负电荷的颗粒表面之间实现静电吸引。原子力显微镜（A FM）测量和二辛可宁酸（BCA）测定结果显示hIgG和hIgG抗体（抗hIgG）之间的结合结构，粒径逐渐增加至152.6 rim（裸）、170.2 nm（hIgG）和178.9 nm（hIgG/抗hIgG）。表面覆盖密度为 331.4 ng/cm(2) (hIgG) 和 320.3 ng/cm(2) (cTnI) 以及 hIgG 与 HyLite-750 标记的 Fab 特异性抗 hIgG 的结合能力（接近 81.2%）表明大部分 hIgG 以 Y 形方向固定，夹心免疫测定结果提供了证据，表明 hIgG 的免疫活性PS纳米颗粒表面的cTnI被保留，因为cTnI-PS纳米颗粒/cTnI（抗原）/检测cTnI-抗体反应的结合活性比用作阴性对照的cTnI-PS纳米颗粒/人血清白蛋白（HSA）/检测cTnI抗体反应的结合活性高5倍。
Anionic sulfate (SO(4)(-))-functionalized polystyrene (PS) nanoparticles were prepared by the thermal decomposion of potassium persulfate (K PS) in the presence of sodium tetra borate via emulsion polymerization. The presence of a SO.: group at a solid/liquid interface of a particle surface was confirmed by a zeta potential value of -40.6 mV as well as the shifting of S 2p spectra toward a lower-binding-energy region around 162.7 eV (2p(3/2)) and 164.4 eV (2p(1/2)) in X-ray photoelectron spectroscopy (X PS) analysis. The electrostatic attraction between positively charged antibodies of human immunoglobulin G (hIgG) and cardiac troponin I (c-In I) and negatively charged particle surfaces was accomplished. The atomic force microscopy (A FM) measurement and bicinchoninic acid (BCA) assay results show binding structure between hIgG and antibodies of hIgG (anti-hIgG) with a gradual increase in particle diameter to 152.6 rim (bare), 170.2 nm (hIgG), and 178.9 nm (hIgG/anti-hIgG). Surface coverage densities of 331.4 ng/cm(2) (hIgG) and 320.3 ng/cm(2) (cTnI) and the binding capacity of hIgG to HyLite-750-labeled Fab-specific anti-hIgG (similar to 81.2%) indicate that the majority of hIgG was immobilized with a Y-shaped orientation, The sandwich immunoassay results provide the evidence that the immunological activity of cTnI on the PS nanoparticle surface was retained because the binding activity of the cTnI-PS nanoparticle/cTnI (antigen)/detection cTnI-antibody reaction showed a 5-fold higher activity than that of the cTnI-PS nanoparticle/human serum albumin (HSA)/detection cTnI antibody used as a negative control.