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功能化荧光核壳纳米颗粒用作白细胞介素-6荧光免疫分析中的荧光标记物。

Functionalized fluorescent core-shell nanoparticles used as a fluorescent labels in fluoroimmunoassay for IL-6.

作者信息

Hun Xu, Zhang Zhujun

机构信息

Department of Chemistry, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an 710062, China.

出版信息

Biosens Bioelectron. 2007 May 15;22(11):2743-8. doi: 10.1016/j.bios.2007.01.022. Epub 2007 Feb 6.

Abstract

Nanoparticle labels conjugated with biomolecules are used in a variety of different assay applications. In this paper, a sensitive fluoroimmunoassay for recombinant human interleukin-6 (IL-6) with the functionalized Rubpy-encapsulated fluorescent core-shell silica nanoparticles labeling technique has been proposed. IL-6 was measured based on the specific interaction between captured IL-6 antigen and functionalized fluorescent core-shell nanoparticles-labeled anti-IL-6 monoclonal antibody. The calibration graph for IL-6 was linear over the range 20-1250 pg ml(-1) with a detection limit of 7 pg ml(-1) (3 sigma). The regression equation of the working curve is I(F)=7.665+32.499[IL-6] (ng ml(-1)) (r=0.9980). The relative standard deviation (R.S.D.) for 11 parallel measurements of 78 pg ml(-1) IL-6 was 3.2%. Furthermore, the application of fluorescence microscopy imaging in the study of the antibody labeling and sandwich fluoroimmunoassay with the functionalized fluorescent core-shell silica nanoparticles was also explored. This proposed method has the advantage of showing the specificity of immunoassay and sensitivity of fluorescent nanoparticle labels technology. The results demonstrate that the method offers potential advantages of sensitivity, simplicity and reproducibility for the determination of IL-6, and is applicable to the determination of IL-6 in serum samples and enabling fluorescence microscopy imaging for the determination of IL-6.

摘要

与生物分子共轭的纳米颗粒标记物被用于各种不同的分析应用中。本文提出了一种采用功能化的钌吡啶配合物包裹的荧光核壳二氧化硅纳米颗粒标记技术检测重组人白细胞介素-6(IL-6)的灵敏荧光免疫分析法。基于捕获的IL-6抗原与功能化荧光核壳纳米颗粒标记的抗IL-6单克隆抗体之间的特异性相互作用来测定IL-6。IL-6的校准曲线在20 - 1250 pg ml⁻¹范围内呈线性,检测限为7 pg ml⁻¹(3σ)。工作曲线的回归方程为I(F)=7.665 + 32.499[IL-6] (ng ml⁻¹)(r = 0.9980)。对78 pg ml⁻¹的IL-6进行11次平行测量的相对标准偏差(R.S.D.)为3.2%。此外,还探索了荧光显微镜成像在功能化荧光核壳二氧化硅纳米颗粒抗体标记和夹心荧光免疫分析研究中的应用。该方法具有免疫分析特异性和荧光纳米颗粒标记技术灵敏度的优点。结果表明,该方法在测定IL-6方面具有灵敏度高、操作简单和重现性好的潜在优势,适用于血清样品中IL-6的测定,并能够通过荧光显微镜成像来测定IL-6。

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