Orouji Afsaneh, Ghamsari Mahdi, Abbasi-Moayed Samira, Akbari Mahmood, Maaza Malik, Hormozi-Nezhad Mohammad Reza
Department of Chemistry, Sharif University of Technology, Tehran, 111559516, Iran.
Department of Analytical Chemistry, Faculty of Chemistry, Kharazmi University, Tehran, 15719-14911, Iran.
Nanoscale. 2025 Mar 28;17(13):7957-7969. doi: 10.1039/d4nr04797d.
The rapid and precise quantification and identification of proteins as key diagnostic biomarkers hold significant promise in allergy testing, disease diagnosis, clinical treatment, and proteomics. This is crucial because alterations in disease-associated genetic information during pathogenesis often result in changes in protein types and levels. Therefore, the design of portable, fast, user-friendly, and affordable sensing platforms rather than a single-sensor-per-analyte strategy for multiplex protein detection is quite consequential. In the present research, a robust multicolorimetric probe based on the inhibited etching of gold nanorods (AuNRs) allowing unambiguous high-performance visual and spectral quantification and identification of proteins in human urine samples was designed. Most recently, we discovered that -bromosuccinimide (NBS) can quickly etch AuNRs with a distinct color change, allowing convenient and accurate visual recognition of all amino acids. Herein, further explorations revealed that the presence of proteins, as amino acids' polymers, reduces the effective concentration of NBS to different amounts and in turn prevents the etching of AuNRs to various degrees, thereby allowing precise quantification and identification of various proteins ranging from phosphatase (ACP), pepsin (Pep), hemoglobin (Hem), and transferrin (TRF) to immunoglobulin G (IgG), lysozyme (Lys), fibrinogen (Fib), and human serum albumin (HSA). The acquired dataset was statistically analyzed using linear discriminant analysis (LDA), partial least-squares regression (PLSR), and hierarchical cluster analysis (HCA) to accurately classify and identify individual proteins and their combinations at various levels. The multivariate regression models indicated that the colorimetric responses were linearly dependent on protein concentrations with low detection limits of around 1 ppm. Most importantly, the proposed multidimensional colorimetric probe was successfully utilized for protein discrimination in real urine samples. The diverse rainbow responses exhibited by the AuNRs in the proposed probe greatly enhance the accuracy of visual detection, making it a practical tool for straightforward protein monitoring in real samples.
将蛋白质作为关键诊断生物标志物进行快速、精确的定量和鉴定，在过敏检测、疾病诊断、临床治疗和蛋白质组学中具有重大前景。这至关重要，因为发病机制中疾病相关遗传信息的改变通常会导致蛋白质类型和水平的变化。因此，设计便携式、快速、用户友好且价格合理的传感平台，而非针对多重蛋白质检测采用单传感器检测一种分析物的策略，具有重要意义。在本研究中，设计了一种基于金纳米棒（AuNRs）抑制蚀刻的强大多比色探针，可对人尿液样本中的蛋白质进行明确的高性能视觉和光谱定量及鉴定。最近，我们发现N-溴代琥珀酰亚胺（NBS）能快速蚀刻AuNRs并伴有明显的颜色变化，从而实现对所有氨基酸的便捷、准确视觉识别。在此，进一步探索发现，作为氨基酸聚合物的蛋白质的存在会将NBS的有效浓度降低到不同程度，进而不同程度地阻止AuNRs的蚀刻，从而能够精确地定量和鉴定从酸性磷酸酶（ACP）、胃蛋白酶（Pep）、血红蛋白（Hem）、转铁蛋白（TRF）到免疫球蛋白G（IgG）、溶菌酶（Lys）、纤维蛋白原（Fib）和人血清白蛋白（HSA）等各种蛋白质。使用线性判别分析（LDA）、偏最小二乘回归（PLSR）和层次聚类分析（HCA）对获取的数据集进行统计分析，以在各个水平上准确分类和鉴定单个蛋白质及其组合。多元回归模型表明，比色响应与蛋白质浓度呈线性相关，检测限低至约1 ppm。最重要的是，所提出的多维比色探针成功用于实际尿液样本中的蛋白质鉴别。所提出的探针中AuNRs呈现的多样彩虹响应极大地提高了视觉检测准确性，使其成为实际样本中直接蛋白质监测的实用工具。
