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在Balb/c小鼠腹水中扩大抗人心肌肌钙蛋白I单克隆抗体的生产及特性鉴定

Scale-up production and characterization of anti-human cardiac troponin I monoclonal antibody in ascitic fluid of balb/c mice.

作者信息

Majid Asiabanha Rezaee, Javad Rasaee Mohammad, Malihe Paknejad, Javad Mohammadnejad

机构信息

a Department of Clinical Biochemistry , Faculty of Medical Sciences, Tarbiat Modares University , Tehran , Iran.

b Department of Biochemistry, Faculty of Medical Sciences , Tehran University of Medical Sciences , Tehran , Iran.

出版信息

J Immunoassay Immunochem. 2017;38(4):389-399. doi: 10.1080/15321819.2016.1274263. Epub 2016 Dec 27.

Abstract

The Human Cardiac Troponin I (hcTnI) is a 210 amino acid protein, 23 kDa in molecular weight. This biomarker is commonly used to diagnose myocardial infarction, micro injury, and acute coronary syndrome (ACS) in patients referring to emergency departments. The American Heart Association (AHA) and European Society of Cardiology (ESC) proposed troponin I as the gold biomarker for early detection of heart attack, especially in myocardial infarction (MI). Therefore, developing monoclonal antibodies against this biomarker could help in for early detection of heart attack. Hybridoma technology is a well-known technique introduced to produce monoclonal antibodies in specialized cells. The aim of this study was to produce large scale of monoclonal antibody against human cardiac troponin I using Hybridoma technology in order to design a diagnostic kit. The monoclonal antibody was produced using conventional Hybridoma technology in ascitic fluid of mouse and characterized for its ability to detect Human Cardiac Troponin I in a rapid test system. The results indicate the successful detection of Troponin I using the obtained monoclonal antibody. According to the achieved results it seems that ascites production of monoclonal antibody is very versatile, inexpensive, and economically useful for monoclonal antibody production.

摘要

人心脏肌钙蛋白I(hcTnI)是一种由210个氨基酸组成的蛋白质,分子量为23 kDa。这种生物标志物常用于诊断急诊科患者的心肌梗死、微损伤和急性冠状动脉综合征(ACS)。美国心脏协会(AHA)和欧洲心脏病学会(ESC)提议将肌钙蛋白I作为早期检测心脏病发作的金标准生物标志物,尤其是在心肌梗死(MI)方面。因此,开发针对这种生物标志物的单克隆抗体有助于早期检测心脏病发作。杂交瘤技术是一种用于在特定细胞中产生单克隆抗体的知名技术。本研究的目的是利用杂交瘤技术大规模生产抗人心脏肌钙蛋白I的单克隆抗体,以设计一种诊断试剂盒。该单克隆抗体采用传统杂交瘤技术在小鼠腹水中产生,并在快速检测系统中对其检测人心脏肌钙蛋白I的能力进行了表征。结果表明,使用获得的单克隆抗体成功检测到了肌钙蛋白I。根据所取得的结果,似乎腹水生产单克隆抗体非常通用、廉价且在经济上对单克隆抗体生产有用。

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