| 消减免疫法制备抗HL60、HL60/ADR表面抗原差异抗体 |
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| 引用本文： | 任思楣,俞云,佘鸣,邵晓枫,师锐赞,彭洪薇,林阳,张秀丽,张砚君,熊冬生. 消减免疫法制备抗HL60、HL60/ADR表面抗原差异抗体[J]. 中国免疫学杂志, 2010, 26(2) |
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| 作者姓名： | 任思楣 俞云 佘鸣 邵晓枫 师锐赞 彭洪薇 林阳 张秀丽 张砚君 熊冬生 |
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| 作者单位： | 1. 中国医学科学院,血液病医院,血液学研究所,实验血液学国家重点实验室,天津,300020
2. 南京中医药大学药学院,南京,210029 |
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| 基金项目： | 国家自然科学基金，天津市科技发展项目 |
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| 摘 要： | 目的:建立可特异识别HL60和HL60/ADR细胞膜表面差异蛋白的抗体库,制备单克隆抗体并研究其生物学活性。方法:通过Cp诱导的消减免疫法免疫BALB/c小鼠,采用传统杂交瘤技术制备可特异识别两种细胞差异膜蛋白的单克隆抗体;采用FACS和激光共聚焦显微镜鉴定其和两种靶细胞结合的特异性和差异性。结果:获得51株候选的差异抗体,成功筛选并纯化其中一株单抗(5F6)。5F6与可特异稳定的识别HL60/ADR细胞,结合率扣除本底为68.2%,而该抗体与HL60细胞的结合率为17%。结论:SI结合差异筛选的方法是制备差异抗体便捷可行的方法,所获得的单克隆抗体与HL60和HL60/ADR细胞膜蛋白结合有特异性和差异性,实验中所获得的单抗是寻找肿瘤耐药的新靶点和发现肿瘤耐药新机制的工具,为研究这些问题开拓了新思路。
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| 关 键 词： | 消减免疫 肿瘤耐药 单克隆抗体 差异识别 |
Production of discrepant monoclonal antibody against HL60 and HL60/ADR by SI technique |
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| REN Si-Mei,YU Yun,SHE Ming,SHAO Xiao-Feng,SHI Rui-Zan,PENG Hong-Wei,LIN Yang,ZHANG Xiu-Li,ZHANG Yan-Jun,XIONG Dong-Sheng. Production of discrepant monoclonal antibody against HL60 and HL60/ADR by SI technique[J]. Chinese Journal of Immunology, 2010, 26(2) |
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| Authors: | REN Si-Mei YU Yun SHE Ming SHAO Xiao-Feng SHI Rui-Zan PENG Hong-Wei LIN Yang ZHANG Xiu-Li ZHANG Yan-Jun XIONG Dong-Sheng |
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| Affiliation: | REN Si-Mei,YU Yun,SHE Ming,SHAO Xiao-Feng,SHI Rui-Zan,PENG Hong-Wei,LIN Yang,ZHANG Xiu-Li,ZHANG Yan-Jun,XIONG Dong-Sheng.State Key Laboratory of Experimental Hematology,Institute of Hematology,CAMS & PUMC,Tianjin 300020,China |
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| Abstract: | Objective:To prepare and characterize specific and discrepant mouse hybridoma antibodies on membrane of HL60 and HL60/ADR cell lines.Methods:BALB/c mice were immunized by subtractive immunization induced Cp(Cyclophosphamide).McAbs were prepared by hybridoma technique,screened and detected by FACS and LSCM.Results:51 candidates and discrepant antibodies were found,and one of them(5F6) was purified and identified.Conclusion:Combination of SI with discrepant screening method should facilitate the preparing and... |
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| Keywords: | Subtractive immunization Tumor drug resistance McAb Discrepant protein |
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