抗人gp130分子单克隆抗体的制备和鉴定

Preparation and characterization of monoclonal antibodies to human gp130

目的 gp130(亦称CD130)可与许多细胞因子受体(IL 6R,IL 11R,OSMR,LIFR,CNTFR和CT 1R)组成受体复合物,并在这类细胞因子的信号转导中起着重要的作用。方法以痘苗病毒为载体构建获得的可溶性CD130(sCD130)为免疫原 ,通过细胞融合的方法 ,得到4株稳定分泌抗CD130单克隆抗体(mAb) (L2,L4a,L4b和L5)的杂交瘤细胞株。结果本组mAb对靶细胞具有识别特异性 ,不但可识别相对分子质量 (Mr)为98000的sCD130分子,也可识别Mr为130000的膜表达型CD130分子(mCD130)。另外 ,mAb还可与CD130分子形成免疫复合物 ,用于免疫沉淀CD130分子 ,但这组抗体中仅mAbL2可部分组断IL 6对靶细胞的增殖作用。结论成功地研制了抗人gp130分子mAb。

Aim To prepare and characterize mAb to human gp130,and the receptor complexes, with an important role in cytokine signal transduction , which consist of gp130 also known as CD130 and cytokine receptors (I1 6R,IL 11R,OSMR,LIFR,CNTFR and CT 1R), Methods Soluble CD130 molecule (SCD130) constructed with vacinia virus as vetor was used as antigen to immunize Balb/c mice. Four hybyidome cell lines(L2,L4a,L4b and L5)that could steadily secret anti gp130 mAb were established. Results The antibody characterization showed that they not only specificaley recognize CD130 molecule with relative molecular mass(Mr)98 000, but also CD130 molecule with Mr 130 000 on the membrane of target cells. Otherwise they could also precipitate CD130 molecule from target cells.The L2 mAb of them could partly block the proliferative effect of IL 6 on target cells. Conclusion Monoclonal antibodies to human gp130 are prepared and characterized. group (P<0.05).The thickness of central cornea in 5 sec group was significant thiner than that of 15 sec group (P<0.05).In 2 d after injury, the area of endothelial wound was(4.13±1.97)mm 2.Some of the wounds had closed completely in 3d after wounding 4 d after injury, all of the corneal wound areas were completely covered by uneven and irregular shaped endothelial cells.In 12 d after injury ,the healed endothelial cells became regular and polygonal.The results indicate that the corneal endothelial injury model by transcorneal freezing in rabbits is reproducable and standardized for the probe′s temperature was accurately maintained with (-120±10)℃ as recorded by the instrument. KEY WORDS trancorneal freezing cornea endothelial injury model rabbit