论文部分内容阅读
目的 应用化学交联法制备抗人成骨肉瘤单克隆抗体 (OSMcAb) 人血清白蛋白(HSA) 甲氨蝶呤 (MTX)偶联物并进行体外细胞毒性实验。方法 采用碳二亚胺、活化酯及SPDP法制备OSMcAb HSA MTX间接偶联物 ;同时应用活化酯法制备出OSMcAb MTX ;并进行体外靶细胞结合实验、细胞毒性实验和集落形成抑制实验。结果 化学交联法制备出的偶联物的性质及药物含量稳定。抗体蛋白活性保存良好。集落形成抑制实验中 ,OSMcAb HSA MTX、OSMcAb MTX的IC5 0 (μg/L)分别为 2 .5 5 0± 0 .114、2 .710± 0 .14 5 ,它们杀伤作用均明显大于游离MTX(IC5 0为 5 .5 10± 1.5 2 9) (P <0 .0 5 )。 2 4h细胞毒性实验中 ,OSMcAb HSA MTX、OSMcAb MTX的对MG 63细胞、T2 4细胞的IC5 0 (μg/L)分别为 66.5 :3 785和 79.8:3 0 2 5 ,偶联物杀伤作用在靶细胞明显强于非靶细胞 (P <0 .0 1)。结论 以化学交联法制备OSMcAb HSA MTX偶联物方法可行 ;偶联物OSMcAb HSA MTX在体外具有良好的抗肿瘤作用。
Objective To prepare anti-human osteosarcoma monoclonal antibody (OSMcAb) human serum albumin (HSA) methotrexate (MTX) conjugate by chemical cross-linking method and to study the cytotoxicity in vitro. Methods OSMcAb HSA MTX indirect conjugate was prepared by carbodiimide, activated ester and SPDP method. At the same time, OSMcAb MTX was prepared by activated ester method. In vitro target cell binding assay, cytotoxicity assay and colony formation inhibition assay were also performed. Results The nature of the conjugate prepared by the chemical cross-linking method and the drug content were stable. Antibody protein activity is well preserved. In colony formation inhibition assay, the IC 50 (μg / L) of OSMcAb HSA MTX and OSMcAb MTX were respectively 2.550 ± 0.114 and 2.710 ± 0.14 5, which were significantly higher than that of free MTX IC 50 was 5.510 ± 1.5 2 9) (P <0.05). In 24 h cytotoxicity assay, IC50 (μg / L) of OSMcAb HSA MTX and OSMcAb MTX against MG 63 cells and T2 4 cells were 66.5: 3 785 and 79.8: 3 025, respectively. The cytotoxicity of the conjugate was The target cells were significantly stronger than non-target cells (P <0.01). Conclusion The preparation of OSMcAb HSA MTX conjugate by chemical cross-linking method is feasible. The conjugate OSMcAb HSA MTX has good antitumor activity in vitro.