Gold nanoparticles-decorated M13 phage SPR probe for dual detection of antigen biomarkers in serum
Gold nanoparticles-decorated M13 phage SPR probe for dual detection of antigen biomarkers in serum
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金纳米颗粒修饰的 M13 噬菌体 SPR 探针用于血清中抗原生物标志物的双重检测
DOI：
10.1016/j.snb.2022.132811
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发表时间：
2022-10-12
影响因子：
8.4
通讯作者：
Zhou, Xin
中科院分区：
文献类型：
--
作者：
Hou, Jinxiu;Xu, Yi;Zhou, Xin
无标记SPR由于其低灵敏度而很少用于真实的样品中的生物标志物检测。在此，我们报告了一种基于噬菌体的SPR（P-SPR）检测未稀释血清中癌胚抗原（CEA）的策略。对M13噬菌体进行基因工程改造，以在pIII蛋白上展示抗CEA单链可变区片段用于抗原靶向，并在pVIII蛋白上展示金结合肽（GBP）用于金纳米颗粒（GNP）结合，以形成GNP修饰的M13噬菌体探针，其可以放大等离子体共振信号。建议的P-SPR方法表现出超灵敏的检测CEA在10分钟内。在理想的条件下，P-SPR表现出异常的灵敏度与检测限（LOD）为0.83 fM，这是约2000倍，传统的SPR和四个数量级更敏感比酶联免疫吸附试验。特别地，所提出的策略可以通过对由P-SPR探针产生的斑块进行裸眼计数来交叉验证SPR结果，从而实现可靠的双重检测系统。此外，P-SPR方案在低于ELISA检测限的浓度下对真实的样品的检测具有优异的传感性能。因此，提出的P-SPR策略具有在液体活检中早期检测肿瘤生物标志物的潜力。
Label-free SPR is rarely used for biomarker detection in real samples due to its low sensitivity. Herein, we report a phage-based SPR (P-SPR) strategy for the detection of carcinoembryonic antigen (CEA) in undiluted serums. M13 phage is genetically engineered to display an anti-CEA single-chain variable region fragment on pIII protein for antigen targeting and to display the gold binding peptide (GBP) at the pVIII protein for gold nanoparticles (GNPs) binding to form the GNPs decorated M13 phage probe that can amplify the plasmon resonance signal. The proposed P-SPR method demonstrated ultrasensitive detection of CEA within 10 min. Under ideal conditions, P-SPR showed exceptional sensitivity with a limit of detection (LOD) of 0.83 fM, which is approximately 2000 times that of the conventional SPR and four orders of magnitude more sensitive than enzyme-linked immunosorbent assay. In particular, the proposed strategy can cross-validate the SPR results through naked-eye counting of the plaques generated by the P-SPR probes, enabling a reliable dual detection system. In addition, P-SPR scheme has excellent sensing performance for the detection of real samples at concentrations lower than LOD of ELISA. Therefore, the proposed P-SPR strategy holds potential for early detection of tumour biomarkers in liquid biopsy.