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	NOK/STYK1单克隆抗体的制备及应用
	张国芳 ; 王银银 ; 常智杰 ; ZHANG Guofang ; WANG Yinyin ; CHANG Zhijie
	2016-03-30 ; 2016-03-30
关键词	酪氨酸蛋白激酶 NOK/STYK1 单克隆抗体 肿瘤 protein tyrosine kinases NOK/STYK1 monoclonal antibody cancer R392
其他题名	Generation and application of the NOK/STYK1 monoclonal antibody
中文摘要	根据NOK/STYK1(Novel Oncogene with Kinase Domain或Serine/Threonine and Tyrosine Receptor Protein Kinase)的结构特点设计并合成抗原,免疫Balb/C小鼠后,取免疫反应最好的小鼠脾脏细胞与骨髓瘤细胞SP2/0进行融合.应用有限稀释法筛选得到能稳定分泌抗NOK/STYK1的单克隆杂交瘤细胞株7G9.该细胞株经过扩大培养后,通过小鼠腹水制备纯化出NOK/STYK1的单克隆抗体.Western blotting和ELISA检测结果显示,纯化出的单克隆抗体能够特异性识别NOK/STYK1,且效价较高.通过免疫荧光标记和免疫组化分析显示,该单克隆抗体能够有效检测多种肿瘤细胞内及恶性肿瘤组织样本中高表达的NOK/STYK1.; Protein tyrosine kinases(PTKs)-mediated signaling transduction pathways play pivotal roles in the regulation of key functions during embryogenesis,development,metabolism and immunity.PTKs enhance the cell proliferation,mobility and viability by activating multiple downstream signaling transduction pathways,whereas abnormal PTKs catalytic activity often leads to oncogenesis.NOK/STYK1(novel oncogene with kinase domain,also known as serine/ threonine and tyrosine receptor protein kinase)is a new member of PTKs that has been identified as another oncogene.However,the mechanism of NOK/STYK1participates in the regulation of signaling transduction and oncogenesis remains elusive.Thus,to generate a specific monoclonal antibody to illustrate the functions of NOK/STYK1is really in need.In this research,we designed and purified some short peptides of NOK/STYK1as antigens to immunize Balb/C mice to obtain the B cells of spleen.Subsequently,we fused the B cells to murine cells SP2/0and then screened and identified the positive hybridoma cells line for production of the monoclonal antibody against NOK/STYK1by ELISA.Fortunately,we found that a hybridoma cell line 7G9has a high production of monoclonal antibody,which was confirmed for the specificity and efficiency by both western blotting and ELISA.This monoclonal antibody was verified to be powerful for detecting the expression of NOK/STYK1 in various cancer cell lines and tissues by both immunefluorescence and immunohistochemical staining respectively.
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://ir.lib.tsinghua.edu.cn/ir/item.do?handle=123456789/147363]
专题	清华大学
推荐引用方式 GB/T 7714	张国芳,王银银,常智杰,等. NOK/STYK1单克隆抗体的制备及应用[J],2016, 2016.
APA	张国芳,王银银,常智杰,ZHANG Guofang,WANG Yinyin,&CHANG Zhijie.(2016).NOK/STYK1单克隆抗体的制备及应用..
MLA	张国芳,et al."NOK/STYK1单克隆抗体的制备及应用".(2016).

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