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bFGF特异性结合噬菌体的活性研究

Activity Study of Specific bFGF-Binding Phage

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【作者】 颜秋霞吴晓萍黄慧贤聂昌君聂燕芳柯实肖健李校堃

【Author】 YAN Qiu-xia1,3 WU Xiao-ping2 HUANG Hui-xian3 NIE Chang-jun3 NIE Yan-fang2KE Shi3 XIAO Jian1 LI Xiao-kun1(1 School of Pharmaceutical Science/Zhejiang Provincial Key Laboratory of Biotechnology PharmaceuticalEngineering Wenzhou Medical College,Wenzhou 325035,China)(2 Institute of Tissue Transplantation and Immunology,Jinan University,Guangzhou 510632,China)(3 Biopharmaceutical Research and Development Center,Jinan University,Guangzhou 510632,China)

【机构】 温州医学院药学院/浙江省生物技术制药工程重点实验室暨南大学医药生物技术研究开发中心暨南大学组织移植与免疫实验中心

【摘要】 研究从噬菌体随机7肽库中筛选的bFGF特异性结合噬菌体的活性。采用ELISA检测3轮淘选后得到的阳性噬菌体克隆对bFGF的结合曲线。通过竞争抑制实验,分析bFGF结合噬菌体与bFGF的竞争结合活性。采用MTT法检测bFGF结合噬菌体对由bFGF诱导的Balb/c3T3细胞增殖的影响。从噬菌体肽库中淘选获得的阳性噬菌体克隆能够特异地与bFGF结合,并呈剂量依赖性,其与固相bFGF的结合能被游离的bFGF竞争抑制,并可抑制由bFGF诱导的Balb/c3T3细胞增殖。bFGF特异性噬菌体可拮抗bFGF的活性,为开发针对bFGF的抗肿瘤短肽类新药提供基础。

【Abstract】 To study the activity of a specific bFGF-binding phage selected from a phage display heptapeptide library.ELISA was applied to assess the bFGF-binding curve of the positive phage clone obtained from 3 rounds screening.The competitive binding activity of the bFGF-binding phage was examined by competitive inhibition assay.The effect of the bFGF-binding phage on the proliferation of Balb/c 3T3 cells induced by bFGF was investigated by MTT method.The positive phage selected from the phage library specifically band to bFGF in a dose-dependent manner,and its binding with the immobilized bFGF was competitively inhibited by free bFGF.The isolated phage could also inhibit the proliferation of Balb/c 3T3 cells induced by bFGF.The bFGF-binding phage could inhibit bFGF activity,which would offer the basis for further developing the anti-tumor short peptide targeting bFGF.

【基金】 浙江省自然科学基金重点项目(Z205755);浙江省卫生高层次创新人才培养工程(李校堃);教育部跨新世纪优秀人才计划(NCET-05-0537)资助项目
  • 【文献出处】 中国生物工程杂志 ,China Biotechnology , 编辑部邮箱 ,2009年01期
  • 【分类号】Q78
  • 【被引频次】2
  • 【下载频次】191
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