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钓取抗人肿瘤坏死因子-α(hTNF-α)抗体Z12重、轻链可变区(VH,VL)基因,根据其氨基酸序列,运用同源模建方法分别模拟VH和VL结构域的空间构象,并搭建Fv片段的整体三维结构,利用分子对接方法建立Fv/hTNF-α作用的复合物模型,据此模型推测Z12抗体识别的表位为hTNF-α141-146位.将hTNF-α分为N端1-91和C端92-157两段进行原核表达,检测结果表明Z12抗体识别的抗原表位位于hTNF-αC端92-157区,初步证实预测结果可靠.进一步的实验研究表明,当把hTNF-α141-146位氨基酸缺失后,Z12抗体识别该缺失体的能力几乎丧失,提示此抗体特异性识别抗原分子141～146位氨基酸残基.
According to the amino acid sequence of the heavy and light chain variable region (VH, VL) of anti-human tumor necrosis factor-α (hTNF-α) antibody, homology modeling was used to simulate the space of VH and VL domains respectively Conformation, and construct the overall three-dimensional structure of Fv fragment, the molecular docking method was used to establish the Fv / hTNF-α complex model, the model speculated that the epitope recognized by the Z12 antibody was hTNF-α141-146 position hTNF-α The results showed that the antigenic epitope recognized by Z12 antibody was located in the region 92-157 of hTNF-αC, and the result of the preliminary confirmation of the prediction was reliable.Further experimental studies showed that, When the hTNF-α141-146 amino acid is deleted, the ability of the Z12 antibody to recognize the deletion is almost lost, suggesting that the antibody specifically recognizes amino acid residues 141-146 of the antigen molecule.