三氧化二砷免疫毫微球的制备及鉴定

 目的制备单抗CD33导向的三氧化二砷免疫蛋白毫微球(CD33-As₂O₃-BSA-NP)并检测其特异性结合APL原代细胞的活性。方法通过N-羟基琥珀酰亚胺基-3-(2-吡基二硫)-丙酸酯(SPDP)交联的方法制备免疫蛋白毫微球。玻片凝集实验、免疫荧光法、光镜和电镜、CD33-As₂O₃-BSA-NP结合的CD33数量测定、检测As₂O₃免疫毫微球的共价连接与活性。结果CD33-As₂O₃-BSA-NP的玻片凝集反应,免疫荧光染色均为阳性;光镜下可看见细胞周围结合微球,电镜下可看见细胞伸出伪足紧密地与免疫毫微球结合在一起;每1g CD33-As₂O₃-BSA-NP表面偶联的CD33抗体数量是14.5 mg。结论制备的CD33-As₂O₃-BSA-NP由共价键连接且特异性的结合APL原代细胞。

Study on Preparation and Determination of Arsenic Trioxide-Loaded Albumin Immuno-nanoparticles

OBJECTIVE To prepare arsenic trioxide-loaded albumin immuno-nanoparticles(As₂O₃-BSA-NP) targeted with Monoclonal Antibody CD33 and test its specific killing effect on APL original cells.METHODS Immuno-nanoparticles were prepared by methods of SPDP crosslinking.Immuno-nanoparticles and its activity were tested by slide agglutination test,immunofluorescent assay,microscopy and scanning electron microscopy,the number of CD33 junctioned the surface of As₂O₃-BSA-NP.RESULTS CD33-As₂O₃-BSA-NP was positive in slide agglutination test and immunofluorescent assay.Under microscopy,the APL original cel1s were rounded with immuno-nanoparticles.The scanning electronmicroscopy revealed the albumin immuno-nanoparticles were tightly connected with the APL original cells.The amount of CD33 juncted with the surface of one gram CD33-As₂O₃-BSA-NP were 14.5 mg.CONCLUSION CD33-As₂O₃-BSA-NP might specifically bind against APL original cel1s by covalent bond.