重组抗人CD22嵌合抗体SM03的^(125)I标记及生物活性  

Bioactivity Assays and Application of ^(125)I Labeled Human Mouse Chimeric Anti-CD22 Monoclonal Antibody SM03

作　　者：路萍萍[1] 孟志云[1] 吴英良[2] 王敏伟[2] 窦桂芳[1] 

机构地区：[1]军事医学科学院野战输血研究所新药临床前评价实验室,北京100850 [2]沈阳药科大学药学院药理系,辽宁沈阳110016

出　　处：《同位素》2008年第1期20-24,共5页Journal of Isotopes

基　　金：北京市科技计划项目(Z0004105040131)

摘　　要：为了探讨125I标记的重组抗人CD22嵌合抗体SM03的生物活性,采用Iodogen法进行单克隆抗体SM03的125I标记,Sephacryl S-300 HR色谱柱分离标记混合物,测定分离后样品的纯度与浓度。采用竞争结合法和酶联免疫吸附法(ELISA)测定纯化后样品的活性。抗体经过放射性标记后,放化纯度>99%,回收率>47%1。25I-SM03的生物活性与未标记抗体差异没有显著性,P=0.907。用125I-SM03筛选CD22抗原表达量最多的细胞,在Raji、Daudi和Ramos细胞中,SM03与Ramos细胞的特异性结合量最多,每个细胞的结合量为1.3 pmol。以上结果表明,Indogen法可以用于单克隆抗体SM03的标记,标记后不影响SM03的生物活性。To investigate the bioactivity and application of levi labeled human mouse chimeric monoclonal SM03, SM03 was labeled with ^125I using Indogen method. The labeled mixture was purified by Sephacryl S-300 HR separation chromospectry. The purity and concentration of separated fractions were determined by HPLC and Protein Assay Kit, respectively. Competitive binding method and ELISA method were used for bioactivity assays. ^125I-SM03 was applied to screen cell lines which express the most abundant CD22 antigen. The purity and recovery of ^125I-SM03 were 〉99% and 〉47%, respectively. The bioactivity of ^125I- SM03 and SM03 hasn＇t significant difference in statistics. Ramos cell line had the strongest 0 special radioactivity when 12s I-SM03 bound with in Raji, Daudi and Ramos cell lines. Indogen method is a good way to label Human mouse chimeric anti-CD22 monoclonal antibody SM03 and the label will not affect the activity of SM03. The ^125I-SM03 not only can be used for detect agent, but also may be put into market for NHL therapy.

关 键 词：^125I标记 人鼠嵌合抗CD22单克隆抗体 SM03 生物活性 

分 类 号：TQ463[化学工程—制药化工] R817[医药卫生—影像医学与核医学]