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为了研究嵌合抗CD20基因工程抗体Fab的抗肿瘤活性及其抗肿瘤机制,本研究采用竞争性免疫荧光抑制实验,证实抗CD20 Fab片段能竞争性抑制亲本鼠源性抗CD20单克隆抗体HI47和Daudi细胞CD20的结合。MTT法测定嵌合抗CD20 Fab对Daudi细胞生长的影响,结果显示嵌合抗CD20 Fab对Daudi细胞的生长具有抑制作用,抑制作用呈剂量依赖性,其IC50为69μg/ml;利用流式细胞仪测定嵌合抗CD20 Fab诱导细胞凋亡作用,结果显示嵌合抗CD20 Fab可诱导 Daudi细胞凋亡,其凋亡率是Rituxan的2倍。这些实验结果证实嵌合抗CD20 Fab通过诱导Daudi细胞凋亡的机制抑制Daudi细胞生长。
In order to study the anti-tumor activity and anti-tumor mechanism of chimeric anti-CD20 genetically engineered Fab, a competitive immunofluorescence inhibition assay was used in this study to confirm that anti-CD20 Fab fragment competitively inhibited the parental murine anti-CD20 monoclonal antibody HI47 Daudi cells CD20 binding. The effect of chimeric anti-CD20 Fab on the growth of Daudi cells was measured by MTT assay. The results showed that chimeric anti-CD20 Fab could inhibit the growth of Daudi cells in a dose-dependent manner with an IC50 of 69 μg / ml. Flow cytometry The effect of chimeric anti-CD20 Fab on apoptosis was evaluated. The results showed that chimeric anti-CD20 Fab induced apoptosis in Daudi cells with a rate of 2-fold higher than that of Rituxan. These experimental results demonstrate that chimeric anti-CD20 Fab inhibits Daudi cell growth by a mechanism that induces apoptosis in Daudi cells.