利用噬菌体表面展示技术筛选转铁蛋白黏附肽的研究

中国生物工程杂志

2010, Vol. 30

Issue (04): 89-94

研究简报

利用噬菌体表面展示技术筛选转铁蛋白黏附肽的研究

卢燕,石屹峰^**

大连工业大学生物与食品工程学院大连 116034

Screening Human Transferrin Binding Peptides by Phage Display Technology

LU Yan,SHI Yi-Feng

School of Biological & Food Engineering, Dalian Polytechnic University, Dalian 116034, China

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摘要：

为了筛选转铁蛋白黏附肽，应用噬菌体表面展示技术经过三轮生物淘选，成功地从随机七肽库中得到黏附转铁蛋白的重组噬菌体克隆，经过相对亲和力常数测定和DNA测序得到4个转铁蛋白黏附肽的序列。实验中以回收率和选择比为操作参数，对淘选进行了优化，并发展了一种基于噬菌体滴度的相对亲和力常数测定方法。转铁蛋白受体是一种有效的肿瘤标记物，利用转铁蛋白为载体可以实现药物靶向运输，因此转铁蛋白黏附肽将是重组蛋白质药物连接转铁蛋白的有用标签。

关键词： 噬菌体表面展示技术; 转铁蛋白; 转铁蛋白黏附肽; 亲和力常数; 蛋白质标签

Abstract:

Phage display technology was used to select transferrin binding peptides (TfB) from a phage fused random heptapeptide library. After three rounds of biopanning, positive recombinant phage clones were enriched. For estimation the efficiency of phage display screening the binding peptides, the selection criteria of recovery yield and specificity ratio in the panning process were developed. The affinity constants of the four most positive TfB clones were assayed with the method of the measurement of phage titres, and the corresponding TfB sequences were obtained through DNA sequencing. Because of transferrin receptor as a tumor biomarker, transferrin has been used as a drug carrier to deliver therapeutics targeting cancer cells. Therefore TfBs will be explored as useful protein tags for binding Tf to improve the transport properties of recombinant protein drugs in vivo.

Key words: Phage display technology Transferrin Transferrin binding peptides Affinity constant Protein tag

收稿日期: 2009-12-21 出版日期: 2010-04-29

基金资助:

辽宁省教育厅重点实验室项目（2008S015）

通讯作者: 石屹峰 E-mail: shiyf@dlpu.edu.cn;

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引用本文:

卢燕石屹峰. 利用噬菌体表面展示技术筛选转铁蛋白黏附肽的研究[J]. 中国生物工程杂志, 2010, 30(04): 89-94.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Y2010/V30/I04/89

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