本文選題：噬菌體抗體 + cyclin��； 參考：《吉林大學(xué)》2006年碩士論文

【摘要】： 細(xì)胞周期調(diào)節(jié)紊亂是導(dǎo)致腫瘤發(fā)生的重要原因,細(xì)胞周期蛋白D1(cyclin D1)作為細(xì)胞進(jìn)入增殖周期后的第一個周期蛋白,在多種腫瘤細(xì)胞中存在過度表達(dá),與腫瘤的發(fā)生,發(fā)展和預(yù)后都有著極為密切的關(guān)系,因此cyclin D1蛋白被認(rèn)為是腫瘤治療中一個重要靶點(diǎn)。 單鏈抗體(single chain variable fragment, scFv)是具有抗原結(jié)合活性的最小的抗體片段,具有分子小,易于穿入組織細(xì)胞,免疫原性小,能通過基因工程技術(shù)大量生產(chǎn)等特點(diǎn),具有較好的應(yīng)用前景,尤其是在腫瘤診斷和治療中具有獨(dú)特的優(yōu)勢。 本研究采用ELISA、Westernblot、Dot-blot、競爭性抑制、親和力測定、免疫熒光染色等實(shí)驗(yàn)方法,對以重組cyclin D1蛋白為抗原,從人源噬菌體抗體庫中篩選出來的抗cyclin D1噬菌體單鏈抗體進(jìn)行了可溶性表達(dá)、純化及活性分析。發(fā)現(xiàn)篩選到的噬菌體單鏈抗體能夠特異性結(jié)合重組cyclin D1蛋白并且具有較強(qiáng)的親和力,同時(shí),噬菌體單鏈抗體還能夠結(jié)合細(xì)胞內(nèi)源性的cyclin D1蛋白。 本實(shí)驗(yàn)獲得的結(jié)果對后續(xù)開展抗cyclin D1胞內(nèi)抗體腫瘤治療的研究奠定了基礎(chǔ)。
[Abstract]:The disorder of cell cycle regulation is an important cause of tumorigenesis. As the first cyclin of cell into the proliferative cycle, cyclin D1(cyclin D1 is overexpressed in many kinds of tumor cells, and it is associated with tumorigenesis. Development and prognosis are closely related, so cyclin D1 protein is considered as an important target in tumor therapy. Single chain variable fragment, scFv) is the smallest antibody fragment with antigen-binding activity. It has the advantages of small molecule, easy penetration into tissue cells, low immunogenicity, and can be produced in large quantities by genetic engineering. Especially in the diagnosis and treatment of cancer has a unique advantage. In this study, the recombinant cyclin D1 protein was used as antigen to express the single chain antibody against cyclin D1 phage selected from human phage antibody library by the methods of competitive inhibition, affinity assay and immunofluorescence staining. Purification and activity analysis. It was found that the screened phage scFv could specifically bind to recombinant cyclin D1 protein and had strong affinity. At the same time, phage scFv could bind to intracellular cyclin D1 protein. The results of this study laid a foundation for the further study of anti-cyclin D 1 anti-tumor therapy.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392;Q78

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