摘要
目的:制备抗抗CD3 ScFv单克隆抗体并研究其生物活性。方法:分离纯化后的抗CD3 ScFv蛋白免疫BALB/c小鼠,采用传统杂交瘤技术制备抗抗CD3 ScFv单克隆抗体;采用ELISA和Western blot鉴定其亚类和抗原结合特异性;采用FACS测定抗抗CD3 ScFv单克隆抗体对Jurkat细胞特异活性。结果:成功筛选出一株能稳定分泌抗抗CD3 ScFv单克隆抗体的杂交瘤细胞株(10B7),其分泌的抗体亚类为IgG1。该抗抗CD3 ScFv单克隆抗体直标后可特异性结合抗CD3 ScFv蛋白和抗CD3抗体。结论:文中所研制的抗抗CD3 ScFv单克隆抗体具有与抗CD3抗体、抗CD3 ScFv蛋白特异结合的活性,在肿瘤导向治疗中抗CD3/抗肿瘤双特异抗体的亲和层析纯化、药代动力学监测等方面具有广泛的应用前景。
Objective：To prepare monoclonal antibody against anti-CD3 ScFv and study its biological activity. Methods：Anti- CD3 ScFv was used for immunizing BALB/c mice. McAb against anti-CD3 ScFv was prepared by hybridoma technique. ELISA and Western blot were used to identify their immunoglobin subtype and specificity. The binding activities of anti-CD3 ScFv McAb against Jurkat cells was detected by FACS. Results：McAb against anti-CD3 ScFv （10B7） was prepared,which belonged to IgG1. It could react with anti-CD3 antibody specifically. Conclusion：McAb against anti-CD3 ScFv has the specific binding activity with anti-CD3 antibody and anti-CD3 ScFv protein, and therefore be potential for tumor target therapy.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2007年第2期142-145,共4页
Chinese Journal of Immunology
基金
国家自然科学基金(批准号30400558
30400405)
天津市科技发展计划项目(批准号05YFGJZJC01200
05YFGPGX06900)资助
