双特异性抗体制备及装配内皮祖细胞可行性研究  

Feasibility study of the bispecific antibody preparation and assembly in endothelial progenitor cells

作　　者：李桂琼[1] 陈庆伟[1] 杨彦[1] 曹广煜[1] 

机构地区：[1]重庆医科大学附属第二医院老年病科,重庆400010

出　　处：《激光杂志》2013年第1期88-89,共2页Laser Journal

基　　金：国家自然科学基金面上项目(Nl:30970826)

摘　　要：目的:通过化学偶联法制备抗肌凝蛋白轻链/CD34双特异性抗体(BsAb),并装配在内皮祖细胞(EPCs)表面。方法:分离和培养大鼠骨髓来源EPCs。通过化学偶联法构建BsAb以及FITC标记的BsAb。分别将10ng、25ng、50ng和100ng的FITC标记的BsAb与1×106个EPCs混合,在荧光显微镜及流式细胞仪下观察抗体与细胞的结合情况,以筛选出抗体装配的最佳浓度。使用上述实验所筛选出的条件,装配BsAb和EPCs备用。结果:通过化学偶联法成功制备抗肌凝蛋白轻链/CD34双特异性抗体(BsAb),并能够装配在内皮祖细胞(EPCs)表面。BsAb在EPCs表面装配的最佳浓度为50ng/106细胞。结论:BsAb构建成功,并在EPCs表面成功装配BsAb。Objective： To, prepare anti - myosin light chaina/CD34 bisific antib〈xty （BsAb） by ehemieal helera）eanjugation, and to assenable them on the surface of EPCs. Methexts： EPCs were extra,ted, isolated and cultmed as describext aove. BsAb and FITC- BsAb were generated by chenfical heterocanjugation a EPCs were tafixeal with FITC- BsAb at lOt,g, 20ng, 50ng and 100ng, the binding aetivity was detected by fluorescence and tlow cylomety to .areen out the optinat concentratian of the antitaty. EPCs were asanflaled with thea BsAbs for further pupose. Results： BsAb was successfully generated and assetadaled on the surface of EPCs. The most an was .50ng/10^6 cells. Conclusions： BsAb was successlizlly generated andther asanlett on the surface of EPCs.

关 键 词：双特异性抗体 内皮祖细胞 化学偶联法 

分 类 号：TN248.1[电子电信—物理电子学]