Preparation and identification of monoclonal antibodies against CD100 molecule]

To prepare and identify the monoclonal antibodies (mAbs) against CD100.BALB/c mice were immunized with eukaryotic expressed CD100 protein. The splenocytes from immunized mice were fused with Sp2/0 cells and the positive hybridoma clones were screened by indirect ELISA and mAbs reacting to CD100 antigen were identified by indirect immunofluorescent staining and flow cytometry with human acute T lymphoblastic leukemia cell line MOLT-4, activated human PBMC, gibbon T lymphocyte cell line MLA-144 and monkey EBV-transformed B lymphoblast cell line(BLCL).Seven hybridomas secreting mAbs against CD100 molecule were obtained. All the mAbs could react to natural CD100 expressed on human MOLT-4 and activated PBMCs as well as monkey BLCL cells, and six out of them could react to gibbon MLA-144 cells.A set of mAbs recognizing CD100 molecule expressed on human, monkey and gibbon cells are obtained, which may provide an useful tool for studying the structure-function relationship of CD100 molecule of human, as well as gibbon and monkey.