Xiaoyong Dai1,
Yaoling Xiong1,
Dandan Xu1,
Linyan Li1,
Zhijian Su2,
Qihao Zhang2,
Qing Zheng1 
For correspondence:- Qing Zheng Email: tzhengq@jnu.edu.cn Tel:+8602085226518
Received: 13 July 2013 Accepted: 2 February 2014 Published: 24 March 2014
Citation: Dai X, Xiong Y, Xu D, Li L, Su Z, Zhang Q, et al. TfR Binding Peptide Screened by Phage Display Technology - Characterization to Target Cancer Cells. Trop J Pharm Res 2014; 13(3):331-338 doi: 10.4314/tjpr.v13i3.3
© 2014 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..
Purpose: To screen an hTfR affinity peptide and investigate its activity in vitro.
Methods: hTfR affinity phage clones were screened from 7-mer phage display library, and their binding ability evaluated by enzyme-linked immunosorbent assay (ELISA). A competitive assay was performed to discover the peptide BP9 (BP9) binding site on the cells. The inhibitory effect of BP9 on the cells was determined using thiazolyl blue (MTT) assay. EGFP-BP9 fusion protein was expressed in E. coli, and its binding and localization on cells were determined by fluorescence microscopy and confocal microscopy, respectively.
Results: After three rounds of panning, recovery efficiency was 48-fold higher than that of the first round. The peptide BP9 sharing 2 identical amino acids to Tf showed high-affinity to hTfR, and possessed strong proliferation inhibition ratio on different tumour cells of 70 % (HepG2 cells)/77 % (SMMC-7221 cells) at a concentration of 0.1 mM, and 85 % (HepG2 cells)/81 % (SMMC-7221 cells) at a concentration of 0.001 mM for 48 h. The recombinant protein EGFP–BP9 could bind to tumour cells and gain entry via the endocytic pathway.
Conclusion: BP9 can bind to TfR and inhibit the proliferation of the tumour cells over-expressing TfR. The DNA sequence coding for BP9 was able to target the macromolecule to combine with TfR. BP9 may possess potential applications in cancer therapy.
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