Aim: To prepare monoclonal antibodies(mAb) against DR5 and characterize their properties.
Methods: BALB/c mice were immunized with DR5, and then mAb was prepared by hybridoma technique. Ig subclass and specificity of mAbs was analyzed by ELISA and flow cytometry, respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA.
Results: Four hybridoma cell lines secreting anti DR5 mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 4 mAbs in ascitic fluid were 1x10(-4) - 5x10(-6). Affinity constant of mAbs were 1x10(9). They recognized 2 different epitopes on DR5 molecule.
Conclusion: Four mAbs against DR5 are prepared successfully, which provides useful reagent for clinical diagnosis and further research.