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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.
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) fusion technique in a conventional way. 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. Ascites titers were between 1: 106 and 1: 108. Western blot analysis of that all the McAbs 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. Conflu sion: Successfully prepared six strains of hybridoma cell secreted monoclonal antibodies against the hexon proteins of adenovirus vector, and provided the foundation foundation of preclinical research of adenovirus vectors.