QCM Biosensor Based on Polydopamine Surface for Real-Time Analysis of the Binding Kinetics of Protein-Protein Interactions

QCM Biosensor Based on Polydopamine Surface for Real-Time Analysis of the Binding Kinetics of Protein-Protein Interactions
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基于聚多巴胺表面的 QCM 生物传感器实时分析蛋白质-蛋白质相互作用的结合动力学

DOI:
10.3390/polym9100482
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发表时间:
2017-10
期刊:
影响因子:
5
通讯作者:
Pei Zhichao
Pei Zhichao
中科院分区:
工程技术3区
文献类型:
--
作者:
Wu Chunli;Li Xueming;Song Siyu;Pei Yuxin;Guo Lili;Pei Zhichao

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研制了一种基于聚多巴胺(PDA)表面的石英晶体微天平(QCM)生物传感器,用于实时分析蛋白质-蛋白质相互作用的结合动力学。通过简单地将金传感器芯片浸入pH为8.5的多巴胺水溶液中,导致PDA膜自发沉积在传感器芯片表面,然后与蛋白质一起培养,通过迈克尔加成和/或希夫碱反应将其固定在PDA涂层的传感器芯片表面,从而制备出生物传感器。本文以单克隆抗肌红蛋白7005抗体(IgG1)与其抗原人心肌肌红蛋白的相互作用为模型系统,实时分析生物传感器表面的生物分子相互作用。在生物传感器表面研究了抗肌红蛋白7005与肌红蛋白相互作用的动力学参数,结果与胺偶联得到的结果一致。基于PDA表面的生物传感器具有良好的可再生性、再现性和特异性。与最常用的胺偶联法在羧化底物上固定蛋白质相比,本文提出的修饰方法简单,温和,不受蛋白质等电点的限制。此外,PDA生物传感器芯片可以很容易地重复使用,这使得QCM生物传感器分析更加高效和经济。
A quartz crystal microbalance (QCM) biosensor based on polydopamine (PDA) surface was developed for real-time analysis of the binding kinetics of protein-protein interactions. The biosensor was fabricated by simply immersing the gold sensor chip into an aqueous dopamine solution at pH 8.5 leading to a spontaneous deposition of PDA film onto the sensor chip surface, which was followed by incubation with the protein to immobilize it onto the PDA-coated sensor chip surface via Michael addition and/or Schiff base reactions. In this paper, the interaction between monoclonal anti-myoglobin 7005 antibody (IgG1) and its antigen human cardiac myoglobin was used as a model system for real-time analysis of biomolecule interactions on the biosensor surface. The kinetic parameters of the interaction between anti-myoglobin 7005 and myoglobin were studied on the biosensor surface, which were consistent with the results obtained via amine coupling. The biosensor based on PDA surface has excellent regenerability, reproducibility, and specificity. Compared with the most frequently/typically used amine coupling method for immobilization of proteins on carboxylated substrates, the modification methodology presented in this paper is simple, mild and is not subjected to the limitations of the isoelectric point (pI) of the protein. In addition, the PDA biosensor chip can be easily reused, which makes QCM biosensor analysis more efficient and cost effective.
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