抗人诱骗受体3多克隆抗体、单克隆抗体的制备及初步应用
发布时间：2018-08-14 13:12
【摘要】： 诱骗受体3(DcR3,又称TR6)是新近发现的一种可溶性肿瘤坏死因子受体(TNFR)超家族成员,可与Fas、HVEM、DR3竞争结合其相应配体FasL、LIGHT及TL1A,但并不介导细胞凋亡,在细胞的生长、分化、死亡以及免疫调节等方面发挥重要作用,与恶性肿瘤、自身免疫疾病及移植排斥等密切相关。 本文将构建的原核表达质粒pET28a(+)/DcR3转化入大肠杆菌(E.coli) BL21中,高效诱导表达并纯化DcR3融合蛋白,纯化并成功复性的蛋白分别免疫家兔和BALB/c小鼠,获得了效价高、特异性强的抗人DcR3多克隆抗体(pAb)和5株单克隆抗体(mAb)。以pAb为包被抗体,以生物素化的mAb为检测抗体,引入生物素-亲合素系统(BAS),建立双抗体夹心BAS-ELISA方法。利用该方法检测到特发性肺纤维化(IPF)、小细胞肺癌及慢性阻塞性肺疾病(COPD)急性加重期患者血清DcR3水平明显高于健康对照。为进一步研究DcR3的功能、应用及探讨其在临床相关疾病发病机制中的作用奠定基础。
[Abstract]:Decoy receptor 3 (DcR3, also known as TR6) is a newly discovered soluble tumor necrosis factor receptor (TNFR) superfamily member. It can compete with Fas, HVEM, DR3 to bind its corresponding ligands FasL, LIGHT and TL1A, but does not mediate cell apoptosis, plays an important role in cell growth, differentiation, death and immune regulation, and plays an important role in malignant tumor, itself. Immune diseases and transplant rejection are closely related.
The prokaryotic expression plasmid pET28a (+) / DcR3 was transformed into E. coli BL21. The fusion protein of DcR3 was highly induced and purified. The purified and renatured proteins were immunized to rabbits and BALB / c mice respectively. The polyclonal antibodies against human DcR3 (pAb) and five monoclonal antibodies (mAb) with high titer and specificity were obtained. The serum levels of DcR3 in patients with idiopathic pulmonary fibrosis (IPF), small cell lung cancer (SCLC) and chronic obstructive pulmonary disease (COPD) at acute exacerbation stage were significantly higher than those in healthy controls. To study the function of DcR3 and lay a foundation for its application in the pathogenesis of clinical related diseases.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R392
本文编号：2182962
[Abstract]:Decoy receptor 3 (DcR3, also known as TR6) is a newly discovered soluble tumor necrosis factor receptor (TNFR) superfamily member. It can compete with Fas, HVEM, DR3 to bind its corresponding ligands FasL, LIGHT and TL1A, but does not mediate cell apoptosis, plays an important role in cell growth, differentiation, death and immune regulation, and plays an important role in malignant tumor, itself. Immune diseases and transplant rejection are closely related.
The prokaryotic expression plasmid pET28a (+) / DcR3 was transformed into E. coli BL21. The fusion protein of DcR3 was highly induced and purified. The purified and renatured proteins were immunized to rabbits and BALB / c mice respectively. The polyclonal antibodies against human DcR3 (pAb) and five monoclonal antibodies (mAb) with high titer and specificity were obtained. The serum levels of DcR3 in patients with idiopathic pulmonary fibrosis (IPF), small cell lung cancer (SCLC) and chronic obstructive pulmonary disease (COPD) at acute exacerbation stage were significantly higher than those in healthy controls. To study the function of DcR3 and lay a foundation for its application in the pathogenesis of clinical related diseases.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R392
【参考文献】
相关期刊论文 前8条
1 李平,黄昌明,张祥福,卢辉山,杨发端,张鲁榕;DcR3和Ki-67在胃癌的表达及临床意义[J];福建医科大学学报;2004年01期
2 钟智强;兰小鹏;杨湘越;李卫滨;张鲁榕;朱忠勇;;DcR3基因在原发性肺癌的扩增和表达及临床意义[J];福州总医院学报;2007年03期
3 陈罡;罗殿中;;原发性肝细胞癌中诱捕受体3的过度表达及临床意义[J];广东医学;2006年10期
4 李连贺;胡俊兰;赵瑞力;岳卓立;;喉癌组织中DcR3蛋白的表达及临床意义[J];临床耳鼻咽喉头颈外科杂志;2007年12期
5 黄志卫,马忠森,王秀丽,吴艳峰,赫国志;外周血单核细胞DcR3基因过度表达在间质性肺疾病发病中的意义[J];中华结核和呼吸杂志;2003年05期
6 吴艳峰,杨俊玲,马忠森,董春玲,秦彦国;DcR_3基因与特发性肺纤维化肺功能的相关性研究[J];中国老年学杂志;2004年10期
7 王秀丽,尹金植,黄志卫,马忠森;特发性肺间质纤维化病人外周血单核细胞中DcR_3基因表达的意义[J];中国老年学杂志;2004年12期
8 ;Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis[J];World Journal of Gastroenterology;2005年38期
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