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Published on: June 23, 2011
[Preparation and characterization of the monoclonal antibody against HIV-1 p24 antigen]
1Department of Virology, Institute of Infectious Diseases, 302th Hospital of PLA, Beijing 100039, China. lba@263.net
Aim:
To establish hybridoma cells secreting monoclonal antibodies (mAbs) against HIV-1 p24 antigen and characterize their properties.
Methods:
BALB/c mice were immunized with purified p24 protein and then the from splenocytes immunized mice were fused with Sp2/0 cells. Monoclonal hybridoma cell lines were obtained by limiting dilution, HAT and HT-selective culture. The specificity of mAbs was identified by Dot blot and indirect ELISA.
Results:
We had established two hybridoma cell lines secreting mAbs against HIV-1 p24 antigen. The titers of two mAbs in ascitic fluid were 1 x 10(-5) and 1.7 x 10(4)-1.8 x 10(4) mol/L, respectively. Both mAbs belonged to IgG1. Indirect ELISA detection demonstrated that both mAbs had no cross-reactivities with other viral antigens, such as HBcAg, HCV RNA positive sera and HIVgp41.
Conclusion:
Two hybridoma cell lines secreting mAbs against HIV-1 p24 antigen have been established successfully, which lays the foundation for detecting HIV-1 p24 antigen.
