抗腺病毒载体单克隆抗体的制备及其初步鉴定

• 作 者: Chunyan Zhang Jinsong Gong Yan
• 机 构: Experimental Medicine Center
• 出 处: 中德临床肿瘤学杂志（英文版）
• 发表日期: 2004年期
• 关键词: gene therapy adenovirus adenovirus vector monoclonal antibodies

Objective: To prepare and identify monoclonal antibodies (McAbs) against the capsid proteins of adenovirus vector. Methods: BALB/c mice were immunized with a mixture of the purified adenovirus vector (Adv) and Al(OH)3. McAbs were produced using cell fusion technique in a conventional way. The sensitivity and specificity of monoclonal antibodies was identified by indirect enzyme linked immunosorbent assay (ELISA), immunocytochemical staining and Western blotting.Results: Six strains of hybridoma cells (A4H11, A8C7, F1H5, G1D2, G4E3 and H2G8) that can stably secrete the IgG1 McAb against Adv were obtained. After 3 months subculture and low concentration of serum adapting culture, six strains retained their stability to secrete McAb. The ascites titers were between 1:106 and 1:108. Western blot analysis demonstrated that all the McAbs reacted with one protein (about 114 kDa) which is present in wild type 3 adenovirus (wtAd3), wild type 5 adenovirus(wtAd5), wild type 7 adenovirus (wtAd7) and adenovirus vector. Conclusion: Successfully prepared six strains of hybridoma cell secreted monoclonal antibodies against the hexon proteins of adenovirus vector, and provided the substantial foundation of preclinical research of adenovirus vectors.