CAJ | 학술논문

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．
근거 NOK/STYK1（Novel Oncogene with Kinase Domain 혹 Serine/Threonine and Tyrosine Receptor Protein Kinase）적결구특점설계병합성항원，면역Balb/C소서후，취면역반응최호적소서비장세포여골수류세포 SP2/0진행융합．응용유한희석법사선득도능은정분비항NOK/STYK1적단극륭잡교류세포주7G9．해세포주경과확대배양후，통과소서복수제비순화출NOK/STYK1적단극륭항체．Western blotting화 ELISA검측결과현시，순화출적단극륭항체능구특이성식별NOK/STYK1，차효개교고．통과면역형광표기화면역조화분석현시，해단극륭항체능구유효검측다충종류세포내급악성종류조직양본중고표체적 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/STYK1 participates in the regulation of signaling transduction and oncogenesis remains elusive.Thus,to generate a specific monoclonal antibody to illustrate the functions of NOK/STYK1 is really in need.In this research, we designed and purified some short peptides of NOK/STYK1 as antigens to immunize Balb/C mice to obtain the B cells of spleen.Subsequently,we fused the B cells to murine cells SP2/0 and then screened and identified the positive hybridoma cells line for production of the monoclonal antibody against NOK/STYK1 by ELISA.Fortunately,we found that a hybridoma cell line 7G9 has 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.