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通过计算机模拟比较十种理论上柔性较好的接头在 ５′ Ｉ Ｌ６ Ｔ Ｎ ＦΔ融合蛋白中对 Ｉ Ｌ６ 和 Ｔ Ｎ ＦΔ空间结构的影响情况，从中选择了 Ｓ Ａ Ｐ Ｇ Ｔ Ｐ接头．以 Ｓ Ａ Ｐ Ｇ Ｔ Ｐ 作为接头的 ５′ Ｉ Ｌ６ Ｓ Ａ Ｐ Ｇ Ｔ Ｐ Ｔ Ｎ ＦΔ和以 Ｐ Ｇ 为接头的５′ Ｉ Ｌ６ Ｐ Ｇ Ｔ Ｎ ＦΔ空间结构预测结果相似． Ｄ Ｎ Ａ 序列分析两种蛋白的接头序列均与设计的一致．５′ Ｉ Ｌ６ Ｓ Ａ Ｐ Ｇ Ｔ Ｐ Ｔ Ｎ ＦΔ和 ５′ Ｉ Ｌ６ Ｐ Ｇ Ｔ Ｎ ＦΔ蛋白的大肠杆菌表达产物经初步分离、纯化及鉴定后，生物学活性及对高表达 Ｉ Ｌ６ 受体肿瘤细胞的杀伤作用比较结果显示：在 Ｌ９２９细胞上，前者的生物学活性是后者的 ２７ 倍；在 Ｕ９３７ 细胞上，前者对肿瘤细胞的抑制率是后者的１３ 倍．它们对高表达 Ｉ Ｌ６ 受体的 Ｕ９３７ 细胞杀伤作用分别是同样突变位点的人 Ｔ Ｎ Ｆα衍生物的３７ 和 ２９ 倍．实验表明， Ｓ Ａ Ｐ Ｇ Ｔ Ｐ作为接头构建的 ５′ Ｉ Ｌ６ Ｓ Ａ Ｐ Ｇ Ｔ Ｐ Ｔ Ｎ ＦΔ融合蛋白优于以 Ｐ Ｇ 作为接头构建的 ５′ Ｉ Ｌ６ Ｐ Ｇ Ｔ Ｎ ＦΔ融合蛋白．
Ten computer simulations were performed to compare the spatial structure of I L6 and T N FΔ in the 5 ’I L6T N FΔ fusion protein with ten flexible connectors in theory. From the selection, the S A P G T P linker . The 5 ’I L6-S A P G T P T N FΔ with S A P G T P as the linker and the 5’ I L6-G G T N F Δ with L G P G as the linker predicted similar results. D N A Sequence Analysis The linker sequences for both proteins are consistent with the design. 5 ’I L6 S PA G T P T N FΔ and 5’ I L6 P G T N FΔ protein of E. coli after preliminary isolation, purification and identification of biological activity and high expression of I L 6 receptor tumor cell killing results compared: the L929 cells, the former biological activity of the latter 2 7 times; in U937 cells, the former inhibition of tumor cells is the latter 1 3 times. Their killing effect on U937 cells with high expression of IL-6 receptors was 3.7 times and 2.9 times that of human TNFα derivatives with the same mutation sites respectively. The experimental results showed that the 5 ’I L6-S A P G T P-T N FΔ fusion protein constructed by S A P G T P as a linker is superior to the 5’ I L6-G G T N FΔ Fusion protein.