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用于蛋白质检测的多功能脱铁铁蛋白纳米颗粒标记物

Versatile apoferritin nanoparticle labels for assay of protein.

作者信息

Liu Guodong, Wang Jun, Wu Hong, Lin Yuehe

机构信息

Pacific Northwest National Laboratory, Richland, Washington 99352, USA.

出版信息

Anal Chem. 2006 Nov 1;78(21):7417-23. doi: 10.1021/ac060653j.

Abstract

A versatile bioassay label based on marker-loaded apoferritin nanoparticles (MLANs) has been developed for sensitive protein detection. Dissociation and reconstitution characteristics at different pH as well as the special cavity structure of apoferritin provides a facile route to prepare nanoparticle labels and avoid the complicated and tedious synthesis process of conventional nanoparticle labels. The optical and electrochemical characteristics of the prepared nanoparticle labels are easily controlled by loading different optical or electrochemical markers. A fluorescence marker (fluorescein anion) and a redox marker [hexacyanoferrate(III)] were used as model markers to load into the cavity of apoferritin nanoparticles for microscopic fluorescence immunoassay and electrochemical immunoassay, respectively. Detection limits of 0.06 (0.39 pM) and 0.08 ng mL(-1) (0.52 pM) IgG were obtained with fluorescein MLAN and hexacyanoferrate MLANs, respectively. The new nanoparticle labels hold great promise for multiplex protein detection (in connection with nanoparticles loaded with different markers) and for enhancing the sensitivity of other bioassays.

摘要

一种基于载有标记物的脱铁铁蛋白纳米颗粒(MLANs)的多功能生物测定标记物已被开发用于灵敏的蛋白质检测。脱铁铁蛋白在不同pH下的解离和重构特性以及其特殊的腔室结构为制备纳米颗粒标记物提供了一条简便途径,避免了传统纳米颗粒标记物复杂繁琐的合成过程。通过加载不同的光学或电化学标记物,可以轻松控制所制备纳米颗粒标记物的光学和电化学特性。分别使用荧光标记物(荧光素阴离子)和氧化还原标记物[六氰合铁(III)酸盐]作为模型标记物加载到脱铁铁蛋白纳米颗粒的腔室中,用于微观荧光免疫测定和电化学免疫测定。荧光素MLAN和六氰合铁MLANs分别获得了0.06(0.39 pM)和0.08 ng mL⁻¹(0.52 pM)的IgG检测限。这种新型纳米颗粒标记物在多重蛋白质检测(与载有不同标记物的纳米颗粒相关)以及提高其他生物测定的灵敏度方面具有巨大潜力。

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