抗Pgp/抗CD3双特异双链抗体的生物学活性研究  

Biological activity of anti-CD3/anti-Pgp diabody

作　　者：高瀛岱[1] 熊冬生[1] 许元富[1] 杨铭[1] 彭晖[1] 邵晓枫[1] 刘汉芝[1] 杨纯正[1] 

机构地区：[1]中国协和医科大学中国医学科学院血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中华微生物学和免疫学杂志》2004年第5期352-355,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家高科技研究发展专项经费资助项目(2 0 0 2AA2Z3 46D)

摘　　要：目的　研究抗Pgp 抗CD3双特异双链抗体介导的特异性靶向杀伤活性。方法　采用亲和层析法分离纯化本室构建的抗Pgp 抗CD3双特异双链抗体可溶性表达产物 ,并用SDS PAGE、Westernblot及抗活细胞间接免疫荧光法鉴定纯化产物 ;采用51 Cr释放试验测定其介导的体外靶向杀伤活性。结果　纯化的抗Pgp 抗CD3双特异双链抗体具有与Jurkat细胞 (CD3+ )和K5 6 2 A0 2 (Pgp+ )细胞结合的活性 ;抗Pgp 抗CD3双特异双链抗体具有介导激活的T淋巴细胞杀伤Pgp表达阳性的耐药肿瘤细胞的作用 ,杀伤作用的强弱显示出效靶比和剂量依赖关系 ,且可被CD3ScFv或PgpScFv特异性的阻断。结论　抗Pgp 抗CD3微型双功能抗体有望成为治疗耐药肿瘤 ,特别是肿瘤残留灶和微小转移灶的治疗的一种新策略。Objective To study the biological activity of anti-CD3/anti-Pgp diabody. Methods The anti-CD3/anti-Pgp diabody was purified by affinity chromatography and analyzed by Western blot and size exclusion chromatography;its antigen-binding activity was examined by FACS. Its biological activity was examined by 51 Cr release test. Results The anti-CD3/anti-Pgp diabody can bind to Jurkat cells (CD3 +) and K562/A02 cells(Pgp +). In the presence of activated human peripheral blood lymphocytes (PBL), the diabody mediates effectively the lysis of the Pgp-overexpressing human leukemia K562/A02, but is much less potent in mediating the lysis of the parent K562. Conclusion The biological activity experiments demonstrated that the diabody the anti-Pgp/anti-CD3 diabody may have great potential in the treatment of MDR cancers.

关 键 词：抗Pgp 抗CD3 双特异双链抗体 生物学活性 基因工程抗体 肿瘤细胞 

分 类 号：R392[医药卫生—免疫学] R318[医药卫生—基础医学]