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国家自然科学基金(30471959)

作品数:3 被引量:5H指数:1
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卵巢癌抗抗独特型单克隆抗体3D12的制备及初步研究被引量:1
2008年
目的:制备卵巢癌抗抗独特型单克隆抗体并进行初步鉴定,为深入研究卵巢癌疫苗的主动免疫机制奠定基础。方法:以卵巢癌抗独特型单克隆抗体6B11作为免疫原,采用杂交瘤技术制备抗抗独特型单克隆抗体3D12。经酶联免疫吸附实验(ELISA)、免疫组织化学、流式细胞学、免疫印迹杂交等检测其特异性结合抗原的性质;竞争抑制实验鉴定其特异性结合的抗原表位。结果:获得1株稳定分泌3D12的杂交瘤细胞,所分泌的3D12能够特异性结合6B11及卵巢癌初始抗原OC166-9,并能竞争抑制COC166-9(抗OC166-9的单克隆抗体Ab1)与OC166-9的结合。免疫印迹杂交显示,3D12与COC166-9免疫沉淀的蛋白的相对分子质量相同。该抗体为IgM,亲和力约为8.34×107L/mol。结论:卵巢癌抗抗独特型单克隆抗体3D12属Ab1样Ab3,间接证实6B11为抗原内影像型抗独特型抗体。
张超崔恒李小平叶雪付天云冯捷昌晓红魏丽惠
关键词:卵巢肿瘤抗体抗独特型抗体单克隆
Humoral immune responses induced by anti-idiotypic antibody fusion protein of 6B11scFv/hGM-CSF in BALB/c mice被引量:4
2006年
Background We have previously developed and characterized a monoclonal anti-idiotype antibody, designated 6B11, which mimics an ovarian carcinoma associated antigen OC166-9 and whose corresponding monoclonal antibody is COC166-9 (Ab1). In this study, we evaluate the humoral immune responses induced by the fusion protein 6B11 single-chain variable fragment (scFv)/human granulocyte macrophage colony-stimulating factor (hGM-CSF) and 6B11scFv in BALB/c mice. Methods The fusion protein 6B11scFv/hGM-CSF was constructed by fusing a recombinant single-chain variable fragment of 6B11scFv to GM-CSF. BALB/c mice were administrated by 6B11scFv/hGM-CSF and 6B11scFv, respectively. Results The fusion protein 6B11scFv/hGM-CSF retained binding to the anti-mouse F(ab)2’ and was also biologically active as measured by proliferation of human GM-CSF dependent cell TF1 in vitro. After immunization with the 6B11scFv/hGM-CSF and 6B11ScFv, BALB/c mice showed significantly enhanced Ab3 antibody responses to 6B11scFv/hGM-CSF compared with the 6B11scFv alone. The level of Ab3 was the highest after the first week and maintained for five weeks after the last immunization. Another booster was given when the Ab3 titer descended, and it would reach to the high level in a week. Conclusion The fusion protein 6B11scFv/hGM-CSF can induce humoral immunity against ovarian carcinoma in vivo. We also provide the theoretical foundation for the application of the fusion protein 6B11scFv/hGM-CSF for active immunotherapy of ovarian cancer.
CHANG Xiao-hong YE Xue CUI Heng FENG Jie LI Yi ZHU Hong-lan YANG Wen-lan FU Tian-yun CHENG Hong-yan GUO Hui-fang
关键词:体液免疫抗体蛋白BALB/C动物实验
卵巢癌抗独特型抗体6B11特异性表位肽在体内诱导抗原特异性体液免疫应答
2005年
目的 探讨卵巢癌抗独特型抗体6B11表位多肽与诱导免疫应答之间的关系。方法 采用化学合成的方法,合成抗独特型抗体6B11轻重链可变区的六条CDR区,分别以三种不同浓度免疫BALB/c小鼠。采用ELISA的方法检测免疫鼠血清,进行体内免疫学功能研究。结果 6B11抗独特型抗体的六条CDR肽分别加入佐剂免疫小鼠后,其中有两条肽可以诱导出Ab3,分别为重链的CDR2区和轻链的CDR2区。结论 这两条肽作为后选肽有可能是6B11抗独特型抗体特异性的CTL表位。
昌晓红崔恒冯捷李巍付天云白符李小平叶雪郭慧方程洪艳成夜霞
关键词:抗独特型抗体卵巢癌表位肽诱导免疫应答CTL表位鼠血清
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