阻断型鼠抗人FL单克隆抗体的制备  

Preparation of Neutralized Mouse Anti-human FL Monoclonal Antibody

作　　者：高文杰[1] 柴青[1] 张学光[1] 居颂光[1] 薛智谋[1] 

机构地区：[1]苏州大学实验动物中心,苏州215123

出　　处：《实验动物与比较医学》2010年第4期293-296,共4页Laboratory Animal and Comparative Medicine

摘　　要：目的研制鼠抗人Flt3-ligand（FL）单克隆抗体（mAb）及其生物学特性研究。方法采用高表达膜FL的基因工程细胞株L929／FL细胞免疫BALB／c小鼠，利用B淋巴细胞杂交瘤技术进行细胞融合，免疫荧光标记和流式细胞术筛选阳性株，采用快速定性试纸法鉴定单抗亚类、间接免疫荧光法检测单抗对肿瘤细胞株的识别谱。然后研究所获得的抗人FL单抗＆FL蛋白对THP-1细胞生长的影响。结果得到稳定分泌鼠抗人FL单克隆抗体的杂交瘤细胞一株，并将其分泌的单克隆抗体命名为3E5。经快速定性试纸法鉴定3E5重链为IgGl，轻链为K链。3E5可有效识别不同肿瘤细胞株表达的FL。FL蛋白能促进THP-1细胞的增殖，加入3E5单克隆抗体后能够抑制这种促进作用。结论获得-阻断型单克隆抗体3E5，为研究FL及Flt3功能提供了实用的基础。Objective To prepare monoclonal antibodies against human FL molecule and analysis of their biological characteristics. Methods The BALB/c mice were immunized with the transfected cell line L929/FL that expressed membrane FL. The splenocytes from the immunized mice were fused with myeloma cell line SP2/0, and then the hybridoma cells were screened with immunostaing and FCM. The isotype of the monoclonal antibody identified by rapid isotyping analysis, and the FL expression profile on tumor cell lines was analyzed by indirect immunofluorescence with mAb 3E5. To analysis the biological function ofmAb 3E5, we added the 3E5 to the RPMI1640 medium containing FL protein to culture THP-1 cells. Results a hybridoma cell line named 3E5 was obtained, which had the property of secreting anti-human FL monoclonal antibody continuously and steadily. Fast-strip method analysis displayed that these mAbs belong to mouse lgG1, K. The mAb 3E5 could recognize FL molecules on different cell lines. FL protein could enhance the proliferation of THP-1, and it could be neutralized by 3E5 mAb. Conclusion Our study established a neutralized anti-human FL mAb which may prove a useful tool for studying the biological function of FL.

关 键 词：FL 杂交瘤 单克隆抗体 

分 类 号：R392-33[医药卫生—免疫学]