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肺癌靶向血栓蛋白hu3D3V_H-tTF的表达及活性鉴定
Expression and activity characterization of thrombosis targeting protein hu3D3V_H-tTF to blood vessel of lung cancer

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肺癌靶向血栓蛋白hu3D3V_H_tTF的表达及活性鉴定.pdf (294.5Kb)
Date
2008
Author
颜江华
王生育
王阶平
王勇军
黄志平
王臻
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  • 医学院-已发表论文 [2663]
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Abstract
目的制备一种用于肺癌血管靶向栓塞治疗的融合蛋白hu3D3VH-tTF,并鉴定其生物学活性。方法利用重叠PCR技术构建tTF与hu3D3VH的融合基因,克隆至表达载体pET22 b(+),在E.coliBL21(DE3)中表达,镍亲和色谱柱纯化目的蛋白。ELISA检测融合蛋白hu3D3VH组分与肺腺癌细胞A549选择性结合活性,凝血实验和FⅩ活化实验鉴定融合蛋白tTF组分的促凝血活性。结果获得序列正确的hu3D3VH/tTF/pET22 b(+)重组子,融合蛋白在E.coliBL21(DE3)中高效表达。纯化后的融合蛋白与肺腺癌细胞A549具有选择性结合活性,并能活化FⅩ、有效促发血液凝固。结论成功构建hu3D3VH/tTF/pET22 b(+)重组子,hu3D3VH/tTF融合蛋白具有hu3D3VH的选择性结合能力同时具有TF的促凝血活性,为开展选择性肺肿瘤血管血栓性栓塞研究奠定了基础。 【英文摘要】 Purpose To prepare the fusion protein of hu3D3VH-tTF for thrombosis targeting therapy of lung cancer and to analyze its biological activities.Methods Fusion gene hu3D3VH-tTF was constructed by overlap PCR,cloned into expression vector pET22 b(+),and expressed in E.coli BL21(DE3).The fusion protein hu3D3VH-tTF was purified through Nickel-affinity chromatography column.The selective binding activity of the hu3D3VH moiety of fusion protein was analyzed using ELISA and the coagulation activity of the tTF moiety...
Citation
中国生化药物杂志 Chinese Journal of Biochemical Pharmaceutics,2008,2991):1-4
URI
https://dspace.xmu.edu.cn/handle/2288/8466

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