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目的获得有效果的抗肝癌基因工程单链免疫细胞因子（ＴＮＦα）。方法将抗肝癌单克隆抗体（ｍＡｂ）ＨＡｂ２５的单链抗体（ｓｃＦｖ２５）与人ＴＮＦα基因偶联，构建抗肝癌免疫细胞因子（ｓｃＦｖ２５－ＴＮＦα），亚克隆人原核ＧＳＴ融合表达载体ｐＧＥＸ ４Ｔ－１中，在大肠杆菌中进行表达。对肝癌细胞ＳＭＭＣ－７７２１涂片进行间接免疫荧光染色，测定纯化后目的蛋白的活性。通过对荷肝癌（ＳＭＭＣ－７７２１）裸鼠进行的初步抑瘤实验，检测ｓｃＦｖ２５－ＴＮＦα的导向治疗效果。结果ｓｃＦｖ２５－ＴＮＦα具有与亲本抗体ＨＡｂ２５类似的抗原结合特异性。 ｓｃＦｖ２５－ＴＮＦα对荷肝癌裸鼠体内直径２ｍｍ左右的肿瘤具有明确的导向杀伤作用，达到３／３完全缓解，明显强于ＴＮＦα对照组，该组有效者只有２／３，且无完全缓解。结论ｓｃＦｖ２５－ＴＮＦα为具有一定效果的抗肝癌双功能抗体。
Objective To obtain effective anti-hepatoma gene engineering single-chain immune cytokines (TNFα). Methods The anti-hepatocarcinoma monoclonal antibody (mAb) HAb25 single chain antibody (scFv25) was conjugated with human TNFα gene to construct an anti-hepatocellular carcinoma immune cytokine (scFv25-TNFα) and subcloned into human prokaryotic GST fusion expression vector pGEX 4T-1. , Expressed in E. coli. Indirect immunofluorescence staining of hepatoma cells SMMC-7721 was performed to determine the activity of the purified protein. Through the preliminary tumor-inhibitory experiments performed on nude mice bearing hepatocellular carcinoma (SMMC-7721), the targeted treatment effect of scFv25-TNFα was examined. Results ScFv25-TNF[alpha] has similar antigen binding specificity as the parent antibody HAb25. The scFv25-TNFα has a clear targeted killing effect on tumors with a diameter of about 2 mm in hepatic cancer-bearing nude mice, reaching a 3/3 complete remission, and is significantly stronger than the TNFα control group. The effective group was only 2/3 and no complete remission was observed. Conclusions scFv25-TNFα is an anti-hepatocarcinoma bispecific antibody with certain effects.