目的探究抗人死亡受体5(dr5)单链抗体(Adr5SCfV)的特异性。方法 ElISA检测Adr5SCfV的效价、亲和力和表位分析。WESTErn blOTTIng检测单链抗体的特异性。MTT检测Adr5SCfV对结肠癌细胞SW480的生长抑制。结果 ElISA显示抗人死亡受体5单链抗体抗体效价为1.2x104。通过间接ElISA证实Adr5SCfV与Adr5MAb识别dr5胞外段(Edr5,系本实验室自己构建)的同一个位点。Adr5SCfV的亲和力常数为2.12x10-2。Edr5与Adr5SCfV能够特异结合。MTT检测结果显示,Adr5SCfV抑制SW480细胞生长呈剂量依赖性,Adr5SCfV终质量浓度分别为0.225 Mg/Ml、0.45 Mg/Ml、0.9 Mg/Ml、1.2 Mg/Ml。200μl时,SW480细胞的存活率分别为97.7%,70.9%、70.1%、23.6%。结论 Adr5SCfV能与Edr5特异性结合,并有较强的活性。The aim of our study is to characterize the property of aDR5ScFv.We employed ELISA to determine the titer,relative affinity and epitope recognition of aDR5ScFv,while Western blotting to analyze the specificity of aDR5ScFv.We also measured the inhibitory effects of aDR5ScFv on colon cancer cell SW480 growth by MTT.We found that the titer of aDR5ScFv was 1.2×104;aDR5ScFv and aDR5mAb recognized the same epitope on DR5 molecule;and the affinity constant of aDR5ScFv was 2.12×10-2.Furthermore,aDR5ScFv could recognize eDR5 specifically and inhibit the growth of SW480 cells in a dose-dependent manner(aDR5ScFv final concentrations of 0.225,0.45,0.9,1.2 mg/ml correspond to SW480 viabilities of 97.7%,70.9%,70.1%,23.6%).Western blotting showed the binding of aDR5ScFv to DR5 was specific.Thus,we concluded that binding of aDR5ScFv to DR5 is specific and aDR5ScFv had strong activity.福建省自然科学基金(C0710046
