SERS与特异性配体的膀胱癌标志物UBC灵敏方法的建立
首发时间:2021-01-14
摘要:膀胱癌是发生在膀胱黏膜上的恶性肿瘤,在全球男性中排第七位,迫切需要基于尿液的生物标志物,用于无创性诊断膀胱癌。尿膀胱癌抗原(Urinary bladder cancer antigen;UBC)作为一种典型的膀胱癌标志物,检测其浓度变化可以作为膀胱癌早期无创诊断的一种方法。目前检测UBC的方法在临床应用中受到一定的限制。在这项研究中采用了噬菌体展示技术为前提筛选出UBC特异性结合肽配体,将该配体与表面增强拉曼光谱(Surface-Enhanced Raman Scattering,SERS)技术结合,基于磁场放大方法将银纳米粒子(AgNPs)作为免疫探针与磁珠粒子多肽探针对UBC抗原进行夹心检测,通过放大的AgNPs表面4-氨基联苯(4-ABP)的拉曼信号,实现了对UBC标准品的定量检测。经筛选获得一条UBC较高亲和性12肽配体,亲和性为KD =4.84×10-7M,UBC检出限LOD = 6.25μg/L,符合临床阈值10μg/L的标准,本研究利用筛选的多肽配体与SERS免疫分析方法结合,有效的对UBC标志物进行定量分析,方法简便、灵敏,为膀胱癌早期诊断提供一种新方法。
关键词: 膀胱癌;UBC检测;SERS
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Establishment of a sensitive method for bladder cancer marker UBC based on SERS and specific ligands
Abstract:Bladder cancer is a malignant tumor that occurs on the bladder mucosa, ranking seventh among men in the world. There is an urgent need for urine-based biomarkers for the non-invasive diagnosis of bladder cancer.Urinary bladder cancer antigen(UBC)is a typical bladder cancer marker, and the detection of its concentration changes can be used as a method for early non-invasive diagnosis of bladder cancer.The current methods for detecting UBC are subject to certain limitations in clinical applications.In this study, phage display technology was used as the premise to screen out UBC-specific binding peptide ligands, and the ligands were combined with Surface-Enhanced Raman Scattering (SERS) technology.Based on the magnetic field amplification method, silver nanoparticles (AgNPs) are used as immune probes and magnetic bead particle peptide probes for sandwich detection of UBC antigens.Through the amplified Raman signal of 4-aminobiphenyl (4-ABP) on the AgNPs surface, the quantitative detection of UBC standards was realized.After screening, a 12-peptide ligand with higher affinity for UBC was obtained. The affinity was KD = 4.84×10-7M, and the detection limit of UBC was LOD = 6.25μg/L, which met the clinical threshold of 10μg/L.In this study, the selected peptide ligands were combined with the SERS immunoassay method to effectively analyze the UBC markers. The method is simple and sensitive, and provides a new method for the early diagnosis of bladder cancer.
Keywords: Bladder Cancer UBC detection SERS
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SERS与特异性配体的膀胱癌标志物UBC灵敏方法的建立
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