[Preparation of a novel functional anti-human CD40 monoclonal antibody and identification of its bioactivity].

AIM

To prepare a novel functional mouse anti-human CD40 monoclonal mAb.

METHODS

Female BALB/c mice of 6-8 weeks old were immunized with CD40 transfectant (L929-CD40) as immunogen. The spleen B cells of the mice were fused with Sp2/0. The hybridoma cells were screened with CD40 transfectant (L929-CD40) by FCM. Fast-strip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio/5C11 competitive assay. The proliferation and cell cycle of tumor cells in vitro were studied by MTT assay and PI staining respectively.

RESULTS

One hybridoma cell line named 2B6 was obtained, which had the property of secreting anti-human CD40 monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 molecule and promoted the proliferation of tumor cells in vitro.

CONCLUSION

One hybridoma cell line which can secret a novel functional mouse anti-human CD40 mAb has been developed successfully. This mAb can specifically recognize human CD40 and influence the growth of tumor cells in vitro.