The establishment and identification of an anti-osteosarcoma anti-body and it′ s cytotoxic effect study / 细胞与分子免疫学杂志

ABSTRACT

AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.