抗人纤维蛋白单抗SZ-63嵌合抗体在小鼠骨髓瘤细胞中的表达研究  

Construction and expression of a functional chimeric murine-human antibody directing against human cross-linked fibrin

作　　者：刘跃[1] 夏利军[1] 傅建新[1] 白霞[1] 万海英[1] 李佩霞[1] 阮长耿[1] 

机构地区：[1]苏州医学院血栓与止血研究室,215007

出　　处：《中华血液学杂志》1997年第9期449-452,共4页Chinese Journal of Hematology

基　　金：核工业总公司核工业科学基金!Y3426292101;江苏省卫生厅科研基金!H9202

摘　　要：目的：减少鼠源性单抗的免疫原性，获得最接近于天然构型的高表达基因工程抗体。方法：将抗人纤维蛋白单抗SZ－63的轻链可变区基因VK和重链可变区基因VH分别与人IgG恒区基因CK、CH进行拼接、扩增，构建了真核表达载体α－lys17－63VK／Hu和α－lys30－63VH／Hu，并应用linofectin方法将其相继导入小鼠骨髓瘤细胞SP2／0中，通过加压筛选获得一抗性细胞克隆，经体外长期传代培养和三次克隆化得到一稳定表达的阳性细胞株。结果：培养上清液中的抗体蛋白表达量为1mg／L，腹水中为40mg／L经Westernblot和竞争实验证实表达的嵌合抗体具有与亲本鼠源单抗相一致的纤维蛋白结合特性。结论：SZ－63嵌合抗体可用于血栓性疾病的导向显像和治疗。Objective: In order to reduce the immunokenicity of the murine antifibrin monoclonal anti body (MAb) SZ-63 to human beings. Methods:First of all, the cDNA sequences encoding the constant re gions of the murine k light chain and γ 1 heavy chain in the SZ-63 MAb were substituted with the corre sponding human genomic sequences. Then the constructed MAb was cloned separately into two selectable expression vectors, and the latter were transfected into mouse myeloma cells (SP2/0) one by one. Re sults :ELISA,Western blot and competition experiment results showed that there was 0. 8~ 1. 0mg/L of chimeric 63IgG in the conditioned medium of selected cell lines,and the expressed IgG had higher antifib rin D-Dimer capability than that of the original MAb. Conclusion: The expressed chimeric antibody can be used as a targeting agent for thrombus imaging and treatment.

关 键 词：纤维蛋白 单克隆抗体 嵌合抗体 血栓性疾病 

分 类 号：R543[医药卫生—心血管疾病] R392.11[医药卫生—内科学]