CAJ | 학술논문

癌症生物标记物是一种新的早期癌症治疗工具,它既可用于癌细胞抑制,也可用于将其杀灭.癌干细胞已被证明是癌症的一个关键标志细胞.最近,随着单克隆抗体技术的发展生物标记物的研究发展迅速,一些重要的癌干细胞标记已被确认.单克隆抗体是癌标记物应用的重要技术,也是临床诊断和治疗重要手段.本研究的癌干细胞标记物CD44,CD133和ABCG2,分别通过Genestar等软件分析,并选定目标抗原特异性的基板的基因,设计出目标基因的模板和引物.经过免疫分析,设计出编码基因,利用PCR克隆技术,将其并插入预先选定的pGEX-4T-1或pET32a质粒上的克隆位点上.载有预定基因的标记物,通过载体转入感受态的E.coli BL21中,在加入IPTG的条件下,诱导其蛋白质的表达,得到目标蛋白质产物.利用SDS-PAGE和Westem Blot analysis,验证其表达的正确性.研究的目标蛋白质片段CD44、CD133和ABCG2,可以用于癌干细胞进一步的研究和其单克隆抗体的制作.
암증생물표기물시일충신적조기암증치료공구,타기가용우암세포억제,야가용우장기살멸.암간세포이피증명시암증적일개관건표지세포.최근,수착단극륭항체기술적발전생물표기물적연구발전신속,일사중요적암간세포표기이피학인.단극륭항체시암표기물응용적중요기술,야시림상진단화치료중요수단.본연구적암간세포표기물CD44,CD133화ABCG2,분별통과Genestar등연건분석,병선정목표항원특이성적기판적기인,설계출목표기인적모판화인물.경과면역분석,설계출편마기인,이용PCR극륭기술,장기병삽입예선선정적pGEX-4T-1혹pET32a질립상적극륭위점상.재유예정기인적표기물,통과재체전입감수태적E.coli BL21중,재가입IPTG적조건하,유도기단백질적표체,득도목표단백질산물.이용SDS-PAGE화Westem Blot analysis,험증기표체적정학성.연구적목표단백질편단CD44、CD133화ABCG2,가이용우암간세포진일보적연구화기단극륭항체적제작.
Cancer biomarkers can provide the chance to develop therapies for cancer during its the natural progression, to either inhibit or eliminate the disease. Cancer stem cells have been proved to be critical in the progression of cancer. Recently,some important cancer stem cells markers have been characterized. The rapid development of cancer biomarker research is attributed to a large extent to the advance of the monoclonal antibody technology. Monoclonal antibodies specific for cancer biomarkers have important clinical use in diagnosis and therapeutics. In the present studies, the cancer stem cell markers,CD44,CD133 and ABCG2 ,were first analyzed for their respective antigenic epitopes by Genestar. Then the gene encoding protein fragment with sound immunogenicity was cloned by PCR and inserted to the multiple cloning site of the pGEX-4T-1 or pET32a plasmid. The resulting expression vector was transformed into the E. coli BL21 strain and the protein expression was induced by the addition of IPTG. Finally,the target protein was characterized by the SDS-PAGE and Western Blot analysis. In further study,these CD44 ,CD133 and ABCG2 target protein fragments will be utilized to develop monoclonal antibodies, which can be used in cancer stem cell research.