消减免疫法制备抗HL60、HL60/ADR表面抗原差异抗体
소감면역법제비항HL60、HL60/ADR표면항원차이항체
Production of discrepant monoclonal antibody against HL60 and HL60/ADR by SI technique
  • 万方数据
    中国免疫学杂志 중국면역학잡지
    CHINESE JOURNAL OF IMMUNOLOGY
    2010年 2期 160-163 ,共4页

    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

    임사미%유운%사명%소효풍%사예찬%팽홍미%림양%장수려%장연군%웅동생

    消减免疫%肿瘤耐药%单克隆抗体%差异识别 消減免疫%腫瘤耐藥%單剋隆抗體%差異識彆
    소감면역%종류내약%단극륭항체%차이식별
    Subtractive immunization%Tumor drug resistance%McAb%Discrepant protein
    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.
    목적:건립가특이식별HL60화HL60/ADR세포막표면차이단백적항체고,제비단극륭항체병연구기생물학활성.방법:통과Cp유도적소감면역법면역BALB/c소서,채용전통잡교류기술제비가특이식별량충세포차이막단백적단극륭항체;채용FACS화격광공취초현미경감정기화량충파세포결합적특이성화차이성.결과:획득51주후선적차이항체,성공사선병순화기중일주단항(5F6).5F6여가특이은정적식별HL60/ADR세포,결합솔구제본저위68.2%,이해항체여HL60세포적결합솔위17%.결론:SI결합차이사선적방법시제비차이항체편첩가행적방법,소획득적단극륭항체여HL60화HL60/ADR세포막단백결합유특이성화차이성,실험중소획득적단항시심조종류내약적신파점화발현종류내약신궤제적공구,위연구저사문제개탁료신사로.
    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 identifying discrepant McAbs for identifying antibodies that can distinguish the differences in proteins expressed in HL60 and HL60/ADR,which is a significative and potential method in the research and target therapy associated drug-resistance.
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