Development of a novel aptasensor using jellyfish collagen as matrix and thrombin detection in blood samples obtained from patients with various neurodisease
Development of a novel aptasensor using jellyfish collagen as matrix and thrombin detection in blood samples obtained from patients with various neurodisease
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DOI：
10.1016/j.snb.2016.01.095
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发表时间：
2016-06-02
影响因子：
8.4
通讯作者：
Emregul, Emel
中科院分区：
文献类型：
--
作者：
Derkus, Burak;Arslan, Yavuz Emre;Emregul, Emel
在本研究中，我们描述了来自根口属水母的胃蛋白酶可溶性去端肽胶原的分离和详细表征以及在凝血酶适体传感中的应用。包括红外光谱、SDS-PAGE 电泳和氨基酸分析在内的各种分析方法已应用于水母胶原蛋白的表征，并与鼠尾胶原蛋白和 BSA 进行比较。当比较来自水母和鼠尾的两种胶原蛋白类型时，观察到水母胶原蛋白含有相对大量的谷氨酸（61个残基/1000个残基）和丙氨酸（63个残基/1000个残基），但脯氨酸含量较低（113个残基/1000个残基）。另一方面，胃蛋白酶可溶的水母胶原蛋白含有少量酪氨酸，表明去端肽胶原蛋白的纯度。电化学阻抗谱是所开发的适体传感器的主要分析技术。所提出的 apta 传感器的检测限为 6.25 nM 凝血酶。使用 apta 传感器和商业 ELISA 试剂盒对多发性硬化症、重症肌无力、癫痫、帕金森病、多发性神经病患者和健康供体的血液和脑脊液样本中的凝血酶水平进行了临床应用分析。结果表明，所提出的系统是凝血酶临床分析的有希望的候选系统。 (C) 2016 Elsevier B.V. 保留所有权利。
In the present study, we describe the isolation and detailed characterization of pepsin-soluble atelocollagen from Rhizostoma pulmo species jellyfish and application towards thrombin apta-sensing. Various analysis methods including infra-red spectroscopy, SDS-PAGE electrophoresis, and amino acid analysis have been applied for the characterization of jellyfish collagen and compared with both rat tail collagen and BSA. When comparing the two collagen types derived from jellyfish and rat tail, jellyfish collagen was observed to contain a relatively high amount of glutamic acid (61 residues/1000 residues) and alanine (63 residues/1000 residues) but low amounts of proline (113 residues/1000 residues). On the other hand, pepsin-soluble jellyfish collagen contained a small quantity of tyrosine indicating the purity of atelo-collagen. Electrochemical impedance spectroscopy is the main analyzing technique of the developed apta-sensor. The proposed apta-sensor has a detection limit of 6.25 nM thrombin. Clinical application were performed with analysis of the thrombin levels in blood and CSF samples obtained from patients with Multiple Sclerosis, Myastenia Gravis, Epilepsy, Parkinson, Polyneuropathy and healthy donors using both the apta-sensor and commercial ELISA kit. The results revealed the proposed system to be a promising candidate for clinical analysis of thrombin. (C) 2016 Elsevier B.V. All rights reserved.