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背景与目的:抗CD20抗体和片段已应用于非霍奇金淋巴瘤的临床治疗,但仍需要开发新的抗CD20抗体和片段(未修饰的或放射性标记的),以治疗对美罗华(利妥昔单抗)无反应的患者。鼠源性抗CD20抗体HI47的嵌合抗体片段Fab和F(ab)’2已被构建。本研究目的是观察HI47(鼠抗-CD20抗体)和其嵌合抗CD20抗体片段抑制肿瘤细胞生长和诱导肿瘤细胞凋亡的作用。方法:用免疫荧光法测定抗CD20抗体与CD20+人B细胞淋巴瘤Raji细胞的结合能力;MTT法测定抗CD20抗体片段对Raji细胞生长的影响;用膜联蛋白Ⅴ染色和DNA琼脂糖凝胶电泳检测和验证抗CD20抗体片段诱导Raji细胞凋亡。结果:HI47和其嵌合的抗CD20抗体片段均可与CD20+Raji细胞结合,结合率可达90%以上;HI47不能与美罗华竞争结合Raji细胞;HI47和其嵌合的抗CD20抗体片段浓度为100μg/ml对Raji细胞的抑制率分别为:(57.0±1.5)%、(65.2±2.5)%、(77.2±3.2)%;单价的抗CD20抗体片段Fab(20μg/ml)能够诱导Raji细胞的凋亡,早期凋亡率为17%。结论:HI47的嵌合抗体片段对Raji细胞有抑制作用,能诱导Raji细胞的凋亡。
BACKGROUND AND AIM: Anti-CD20 antibodies and fragments have been used in the clinical treatment of non-Hodgkin’s lymphoma. However, there is still a need to develop new anti-CD20 antibodies and fragments (unmodified or radiolabeled) for the treatment of rituximab Fluoxetine) non-responsive patients. Chimeric antibody fragments Fab and F (ab) ’2 of murine anti-CD20 antibody HI47 have been constructed. The purpose of this study was to observe the effect of HI47 (murine anti-CD20 antibody) and its chimeric anti-CD20 antibody fragment on tumor cell growth inhibition and tumor cell apoptosis induction. Methods: The binding ability of anti-CD20 antibody to CD20 + human B cell lymphoma Raji cells was measured by immunofluorescence. The effect of anti-CD20 antibody fragment on the growth of Raji cells was determined by MTT assay. The expression of anti-CD20 antibody was detected by Annexin V staining and DNA agarose gel electrophoresis Detection and Validation of Anti-CD20 Antibody Fragment Induce Raji Cell Apoptosis. Results: Both HI47 and its chimeric anti-CD20 antibody fragments could bind to CD20 + Raji cells with a binding rate of more than 90%. HI47 could not compete with Rituximab for binding to Raji cells. The concentration of HI47 and its chimeric anti-CD20 antibody fragment was The inhibitory rates of 100μg / ml Raji cells were (57.0 ± 1.5)%, (65.2 ± 2.5)% and (77.2 ± 3.2)% respectively. Monovalent anti-CD20 antibody fragment Fab Apoptosis, early apoptosis rate was 17%. Conclusion: The chimeric antibody fragment of HI47 can inhibit Raji cells and induce apoptosis of Raji cells.