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利用 MnO-PEI/Ni/Au/适配体作为新型纳米马达从人血清样本中分离 HL-60 癌细胞,并通过适配体/金纳米粒子-聚(3,4-亚乙基二氧噻吩)修饰的 GC 电极进行电化学测定。

Isolation of HL-60 cancer cells from the human serum sample using MnO-PEI/Ni/Au/aptamer as a novel nanomotor and electrochemical determination of thereof by aptamer/gold nanoparticles-poly(3,4-ethylene dioxythiophene) modified GC electrode.

机构信息

Nano Drug Delivery Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran; Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Department of Chemistry, Razi University, Kermanshah, Iran.

出版信息

Biosens Bioelectron. 2018 Jul 1;110:141-146. doi: 10.1016/j.bios.2018.03.034. Epub 2018 Mar 17.

Abstract

Herein, aptamer-modified self-propelled nanomotors were used for transportation of human promyelocytic leukemia cells (HL-60) from a human serum sample. For this purpose, the fabricated manganese oxide nanosheets-polyethyleneimine decorated with nickel/gold nanoparticles (MnO-PEI/Ni/Au) as nanomotors were added to a vial containing thiolated aptamer KH1C12 solution as a capture aptamer to attach to the gold nanoparticles on the surface of nanomotors covalently. The aptamer-modified self-propelled nanomotors (aptamer/nanomotors) were then separated by placing the vial in a magnetic stand. The aptamer-modified self-propelled nanomotors were rinsed three times with water to remove the non-attached aptamers. Then, the resulting aptamer/nanomotors were applied for the on-the-fly" transporting of HL-60 cancer cell from a human serum sample. To release of the captured HL-60 cancer cells, the complementary nucleotide sequences of KH1C12 aptamer solution (releasing aptamer) that has a with capture aptamer was added to phosphate buffer solution (1 M, pH 7.4) containing HL-60/aptamer/nanomotors. Because of the high affinity of capture aptamer to complementary nucleotide sequences of aptamer, the HL-60 cancer cells released on the surface of aptamer/nanomotors into the solution. The second goal of the present work was determining the concentration of HL-60 cancer cell in the human serum samples. The electrochemical impedance spectroscopy technique (EIS) was used for the determination of HL-60 cancer cell. The concentration of separated cancer cell was determined by aptamer/gold nanoparticles-poly(3,4-ethylene dioxythiophene) modified GC electrode (GC/PEDOT-Au/aptamer ). The proposed aptasensor exhibited a good response to the concentration of HL-60 cancer cells in the range of 2.5 × 10 to 5 × 10 cells mL with a low limit of detection of 250 cells mL.

摘要

在此,我们使用适配体修饰的自推进纳米马达将人早幼粒细胞白血病细胞(HL-60)从人血清样本中运输出来。为此,将制备的氧化锰纳米片-聚乙烯亚胺修饰的镍/金纳米粒子(MnO-PEI/Ni/Au)作为纳米马达加入含有巯基化适配体 KH1C12 溶液的小瓶中,作为捕获适配体通过共价键附着在纳米马达表面的金纳米粒子上。然后,将含有纳米马达的小瓶置于磁场中,分离出适配体修饰的自推进纳米马达(适配体/纳米马达)。用去离子水将适配体修饰的自推进纳米马达冲洗三次,以去除未附着的适配体。然后,将得到的适配体/纳米马达用于从人血清样本中“实时”运输 HL-60 癌细胞。为了释放捕获的 HL-60 癌细胞,将 KH1C12 适配体溶液(释放适配体)的互补核苷酸序列添加到含有 HL-60/适配体/纳米马达的磷酸盐缓冲溶液(1M,pH7.4)中。由于捕获适配体与适配体的互补核苷酸序列具有高亲和力,因此将 HL-60 癌细胞从适配体/纳米马达表面释放到溶液中。本工作的第二个目标是确定人血清样品中 HL-60 癌细胞的浓度。采用电化学阻抗谱技术(EIS)测定 HL-60 癌细胞。通过适配体修饰的金纳米粒子-聚(3,4-亚乙二氧基噻吩)修饰的 GC 电极(GC/PEDOT-Au/aptamer)测定分离癌细胞的浓度。所提出的适配体传感器对 HL-60 癌细胞浓度在 2.5×10 到 5×10 细胞 mL 范围内表现出良好的响应,检测限低至 250 个细胞 mL。

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