CAJ | 학술논문

目的:鼠抗人CD83功能性单克隆抗体(mAb)的研制及其生物学特性的鉴定.方法:以人CD83转基因细胞L929/CD83为免疫原,常规免疫6～8周龄的雌性BALB/c小鼠;采用B淋巴细胞融合技术,将免疫小鼠脾脏细胞与Sp2/0融合,以L929/CD83、293/CD83转基因细胞为抗体筛选阳性细胞,经免疫荧光标记分析对杂交瘤进行反复筛选和多次的克隆化培养;采用Ig类和亚类快速定性试纸法、染色体核型分析、竞争结合抑制试验、间接免疫荧光法、Western blot对mAb的生物学特性进行鉴定.结果:获得1株稳定分泌鼠抗人CD83mAb的杂交瘤细胞株(命名为9D8),该mAb能特异性地识别成熟的DC、活化的T细胞及肿瘤细胞株Daudi、8226表达的CD83分子,该mAb识别的位点不同于商品化抗人CD83mAb(HB15e).结论:成功地研制1株鼠抗人CD83mAb,识别位点与HB15e不同,为更好地研究该分子的功能提供良好的物质基础.
목적:서항인CD83공능성단극륭항체(mAb)적연제급기생물학특성적감정.방법:이인CD83전기인세포L929/CD83위면역원,상규면역6～8주령적자성BALB/c소서;채용B림파세포융합기술,장면역소서비장세포여Sp2/0융합,이L929/CD83、293/CD83전기인세포위항체사선양성세포,경면역형광표기분석대잡교류진행반복사선화다차적극륭화배양;채용Ig류화아류쾌속정성시지법、염색체핵형분석、경쟁결합억제시험、간접면역형광법、Western blot대mAb적생물학특성진행감정.결과:획득1주은정분비서항인CD83mAb적잡교류세포주(명명위9D8),해mAb능특이성지식별성숙적DC、활화적T세포급종류세포주Daudi、8226표체적CD83분자,해mAb식별적위점불동우상품화항인CD83mAb(HB15e).결론:성공지연제1주서항인CD83mAb,식별위점여HB15e불동,위경호지연구해분자적공능제공량호적물질기출.
AIM: To prepare and characterize a novel anti-human CD83 monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD83 transfectant (L929/CD83) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells. The hybridoma cells were screened with CD83 transfectant (L929/CD83 and 293/CD83) by FCM. The biological characteristics of antibody were investigated by rapid isotyping analysis, karyotype analysis, competitive inhibition test and Western blot. RESULTS: One hybridoma cell line named 9D8 was obtained, which had the property of secreting antihuman CD83 monoclonal antibody steadily, This mAb specifically recognized CD83 molecule expressed on human mature DC, activated T cells, and tumor cell line Daudi, myeloma cell line 8226. This mAb can recognize a distinct epitope from comercial mAb(HB15e). CONCLUSION: One hybndoma cell line has been developed successfully, which may lay a fundation for further study on the function of this molecule.