本文用TPA和LPS刺激培养的HL6 0细胞作为靶的 ,通过胞内染色技术和流式细胞仪分析了抗TNF α单链抗体 (ScFv)的活性。结果表明 ,抗TNF αScFv和其亲本单克隆抗体 (mAb)与TNF α竞争作用显示平均荧光强度 (MFC)为 2 2 7% ,而mAb与TNF α结合显示MFC为 6 9 5 % ,抗TNF αScFv E tag与TNF α结合显示MFC为 48 8% ,空白和阴性对照的MFC分别为1 1%和 2 5 %。相对于mAb ,ScFv的抑制率是 78 37% ,抗原相对结合力是 6 8 17%。本文的结果说明抗TNF αScFv具有与其亲本单抗相同表位的结合活性 ,也为定量和直观测定ScFv的活性提供了实验依据。 In this study, cultured HL60 cells were treated with TPA and LPS as targets. The activity of anti-TNFα single chain antibody (ScFv) was analyzed by intracellular staining and flow cytometry. The results showed that the anti-TNFαScFv and its parent monoclonal antibody (mAb) competed with TNFα to show an average fluorescence intensity (MFC) of 22.7%, while binding of mAb to TNFα showed that the MFC was 695% and the anti-TNFαScFv Binding of E-tag to TNFα showed a MFC of 48.8%, with blank and negative controls of 11% and 25%, respectively. Compared with the mAb, the inhibition rate of ScFv was 78 37%, and the relative binding antigen was 6 8 17%. Our results demonstrate that anti-TNFαScFv has the same binding activity as its parent mAb and provides an experimental basis for the quantitative and visualization of ScFv activity.