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生物素衍生金纳米粒子网络作为 C 反应蛋白免疫测定的标记物。

Nets of biotin-derived gold nanoparticles as a label for the C-reactive protein immunoassay.

机构信息

Saratov State University, 83 Astrakhanskaya Street, Saratov, 410012, Russia.

出版信息

Anal Bioanal Chem. 2021 Nov;413(27):6867-6875. doi: 10.1007/s00216-021-03645-5. Epub 2021 Sep 24.

Abstract

This study presents a promising approach for the one-pot generation of the biotin-derived gold nanoparticles (GNPs@biotin). We report a direct method for the reduction of tetrachloroauric acid with biotin and generation of the labels due to nets formed via biotin-streptavidin interactions. The synthesized GNPs@biotin have a characteristic plasmon maximum, excellent colloidal stability, and streptavidin coupling efficiency. The size of the GNPs@biotin:streptavidin nets and the efficiency of interaction with specific antibodies can be easily customized by the component concentrations and time of their interaction. Moreover, the proposed labels require no additional reagents or manipulations for the synthesis, separation, or purification. The developed labels were applied for the detection of the model antigen of C-reactive protein (CRP) as a major inflammation biomarker. The assembling labels demonstrated a competitive advantage limit of CRP detection (LOD) of 1.2 ng/mL and a limit of quantification (LOQ) of 3.9 ng/mL in human plasma comparable to classical immunoassays. Moreover, the proposed approach is universal and can be potentially applied for the quantitative determination of other biomarkers in a variety of immunoassays in a combination with specific biotinylated antibodies.

摘要

本研究提出了一种很有前途的方法,可用于一锅法生成生物素衍生的金纳米粒子(GNPs@biotin)。我们报告了一种直接用生物素还原四氯金酸并生成标记物的方法,这是由于生物素-链霉亲和素相互作用形成的网络所致。合成的 GNPs@biotin 具有特征等离子体最大值、出色的胶体稳定性和链霉亲和素偶联效率。GNPs@biotin:链霉亲和素网络的大小和与特定抗体相互作用的效率可以通过其相互作用的组分浓度和时间轻松定制。此外,所提出的标记物在合成、分离或纯化过程中无需额外的试剂或操作。所开发的标记物用于检测 C 反应蛋白(CRP)作为主要炎症生物标志物的模型抗原。组装标记物在人血浆中的 CRP 检测(LOD)的竞争优势限制为 1.2ng/mL,定量检测(LOQ)的限制为 3.9ng/mL,与经典免疫分析相当。此外,该方法具有通用性,并且可以与特异性生物素化抗体结合,潜在地应用于各种免疫分析中其他生物标志物的定量测定。

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