Department of Applied Chemistry, Dong-Eui University, Busan 47340, Republic of Korea.
Department of Applied Chemistry, Dong-Eui University, Busan 47340, Republic of Korea.
J Immunol Methods. 2024 Apr;527:113648. doi: 10.1016/j.jim.2024.113648. Epub 2024 Feb 18.
Tumor necrosis factor-alpha, TNF-α, a cytokine recognized as a key regulator of inflammatory responses, is primarily produced by activated monocytes and macrophages. Measuring TNF-α levels serves as a valuable indicator for tracking several diseases and pathological states. Gold nanotechnology has been identified as a highly effective catalyst with unique properties for measuring inflammatory cytokines. This study aimed to synthesize gold nanoclusters (AuNCs) and the AuNCs-streptavidin system, along with their characterizations and spherical morphology. The detection of TNF-α antigen with AuNCs was determined, and a new immunoassay-based AuNCs analytical platform was studied. In this study, it was demonstrated that the synthesized AuNCs and AuNCs-streptavidin showed a bright-yellow appearance with absorption peaks at A and A nm, respectively. The approximately spherical shape was observed by TEM analysis. The AuNCs demonstrated a sensitivity limit for the detection of the TNF-α antigen, with a linear dose-dependent detection range of less than 1.25 ng/mL. The products of the band sizes and band intensities were proportional to the amount of TNF-α in the range of ∼80 kDa, ∼55 kDa, and ∼ 25 kDa in western blot analysis. The TNF-α in cell lysate was successfully detected using an immunoassay after the activation of RAW264.7 cells with lipopolysaccharide (LPS). This assay may serve as a viable alternative for TNF-α detection with high speed, sensitivity, and qualities, ensuring its broad applications.
肿瘤坏死因子-α（TNF-α），一种被认为是炎症反应关键调节因子的细胞因子，主要由激活的单核细胞和巨噬细胞产生。测量 TNF-α 水平可作为跟踪多种疾病和病理状态的有价值指标。金纳米技术已被确定为一种具有独特性质的高效催化剂，可用于测量炎症细胞因子。本研究旨在合成金纳米簇（AuNCs）和 AuNCs-链霉亲和素系统，并对其进行特性和球形形态的研究。通过 AuNCs 检测 TNF-α 抗原，并研究了一种基于免疫测定的新型 AuNCs 分析平台。在这项研究中，证明了合成的 AuNCs 和 AuNCs-链霉亲和素分别具有 A 和 A nm 的吸收峰，呈现明亮的黄色外观。TEM 分析表明其具有近似球形的形态。AuNCs 对 TNF-α 抗原的检测具有灵敏度限制，检测范围的线性剂量依赖性小于 1.25ng/mL。条带大小和强度的产物与 Western blot 分析中约 80 kDa、约 55 kDa 和约 25 kDa 范围内 TNF-α 的量成正比。在用脂多糖（LPS）激活 RAW264.7 细胞后，通过免疫测定成功检测到细胞裂解物中的 TNF-α。这种测定方法可能是一种具有高速、高灵敏度和高质量的 TNF-α 检测的可行替代方法，确保了其广泛的应用。
