摘要
建立抗人成骨肉瘤单克隆抗体杂交瘤并对杂交瘤细胞及抗体特性进行研究．方法：利用我室自建的成骨肉瘤细胞系（OS－9607）免疫BALB八小鼠，取其脾细胞与骨髓瘤细胞SPZ／O融合，经筛选和克隆化，制备杂交瘤细胞系，用免疫组化ABC方法研究杂交瘤细胞分泌的单克隆抗体特性．结果：制得2株能持续、稳定分泌抗体的杂交瘤细胞系．其中6E7和5D3株杂交瘤细胞经过100d连续培养，单克隆抗体6E7mAb和5D3mAb分泌稳定、特异性强、效价高．免疫组化检测结果发现，6E7mAb和5D3mAb相应抗原在成骨肉瘤组织及成骨肉瘤细胞系呈高度表达（SO％），1O种正常组织中未见阳性反应；中度以上梁色者占阳性总数的80％以上，6E7mAb主要分布于细胞核上，5D3mAb主要分布于细胞质内．结论：此2株杂交瘤细胞分泌的单克隆抗体具有成骨肉瘤细胞特异亲合性，它们的成功研制为发现新的成骨肉瘤生化标志、成骨肉瘤的早期诊断及免疫治疗提供了可能性．
AIM: To establish anti-osteosarcoma antibodyproducing hybridoma cell lines and to study the characteriza-tion of the monoclonal antibodies. METHODS: BALB/c micewere immunized with human osteosarcoma cells OS-96o7 andthe immunized spleen cells were fused with SP2/O cells toraise hybridomas after inoculation for three times. The posi-tive clones of hybridoma were screened with ELISA and im-munochemistry methods using OS-96O7 and normal hepato-cellular cells seeded in 96-well plates and antibodies from thepositive clones were further identified with the immunohis-tochemistry method. RESULTS: Two hybridoma cell lineswere established and both of them, 6E7 and 5D3 secretedhigh quality mAbs steadily. 6E7 and 5D3 cells had all thecharacteristics of hybridoma. The mAbs' corresponding anti-gens were specifica1ly and highly expressed in human os-teosarcoma- With enzyme-labe1ed immunohistochemicalstaining on formaldehyde-fixed sections from human osteosar-coma, it was found that more than 8O percent of the speci-mens expressed the corresponding antigen- No positive ex-pression was observed in 1O kinds of normal tissues. CON-CLUSION: Two high quality hybridomas are cultured and themAbs secreted by the two hybridomas have osteosarcomaspecificity. Further research is needed to characterize the bio-chemical properties of the mAb corresponding antigens and todefine the clinical usefulness of the mAbs in either the diag-nosis or the treatment of osteosarcoma.
出处
《第四军医大学学报》
1999年第12期1045-1047,共3页
Journal of the Fourth Military Medical University
关键词
成骨肉瘤
杂交瘤
单克隆抗体
osteosarcoma
hybridoma
antibodies, monoclonal
immunohistochemistry
