Analytical and Bioanalytical Chemistry

, Volume 410, Issue 3, pp 1071–1077 | Cite as

Selection and identification of transferrin receptor-specific peptides as recognition probes for cancer cells

  • Yuyu Tan
  • Wenli Liu
  • Zhi Zhu
  • Lijun Lang
  • Junxia Wang
  • Mengjiao Huang
  • Mingxia Zhang
  • Chaoyong Yang
Research Paper
Part of the following topical collections:
  1. ABCs 16th Anniversary


Since the transferrin receptor (CD71 or TFRC) is known to be highly expressed in numerous cancers, CD71 has become an attractive target in cancer research. Acquiring specific molecular probes for CD71, such as small molecular ligands, aptamers, peptides, or antibodies, is of great importance for cancer cell recognition and capture. In this work, we chose CD71 as the target for phage display, and after four rounds of positive selection and one round of negative selection, the specific phage library was enriched. After verification and sequence analysis, six peptides were identified to be able to bind to CD71 with high specificity. The specific recognition of the CD71-positive cells was confirmed by flow cytometry and confocal microscopy. Competition experiments demonstrated that peptide Y1 and transferrin (TF) were bound to distinct sites on CD71, indicating that peptide Y1 could replace TF as a potential probe for cell imaging and drug delivery, thus avoiding competition by endogenous TF and side effects.

Graphical abstract

Six peptides were successfully isolated using in vitro biopanning against CD71 with high specificity and affinity. Peptides Y1 and Y2 would be powerful tools in biosensors and biomedicine due to their unique properties.


Phage display Biopanning CD71 Cancer cell Imaging 



We thank the National Science Foundation of China (81602206, 21325522, 21422506, 21435004, 21521004), National Basic Research Program of China (2013CB933703), Program for Changjiang Scholars and Innovative Research Teams in University (IRT13036), National Found for Fostering Talents of Basic Science (NFFTBS, J1310024), and China Postdoctoral Science Foundation (2016M592089) for their financial support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2017_664_MOESM1_ESM.pdf (670 kb)
ESM 1 (PDF 669 kb)


  1. 1.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61(2):69–90.CrossRefGoogle Scholar
  2. 2.
    Oken MM, Creech RH, Tormey DC, Horton J, Davis TE, McFadden ET, et al. Toxicity and response criteria of the Eastern Cooperative Oncology Group. Am J Clin Oncol Cancer. 1982;5(6):649–55.CrossRefGoogle Scholar
  3. 3.
    Ferrari M. Cancer nanotechnology: opportunities and challenges. Nat Rev Cancer. 2005;5(3):161–71.CrossRefGoogle Scholar
  4. 4.
    Allen TM, Cullis PR. Drug delivery systems: entering the mainstream. Science. 2004;303(5665):1818–22.CrossRefGoogle Scholar
  5. 5.
    Mo Z-C, Ren K, Liu X, Tang Z-L, Yi G-H. A high-density lipoprotein-mediated drug delivery system. Adv Drug Deliv Rev. 2016;106:132–47.CrossRefGoogle Scholar
  6. 6.
    Neves AR, Queiroz JF, Reis S. Brain-targeted delivery of resveratrol using solid lipid nanoparticles functionalized with apolipoprotein E. J Nanobiotechnol. 2016;14:27–43.CrossRefGoogle Scholar
  7. 7.
    Pan WH, Kastin AJ, Zankel TC, van Kerkhof P, Terasaki T, Bu GJ. Efficient transfer of receptor-associated protein (RAP) across the blood-brain barrier. J Cell Sci. 2004;117(21):5071–8.CrossRefGoogle Scholar
  8. 8.
    Salvati A, Pitek AS, Monopoli MP, Prapainop K, Bombelli FB, Hristov DR, et al. Transferrin-functionalized nanoparticles lose their targeting capabilities when a biomolecule corona adsorbs on the surface. Nat Nanotechnol. 2013;8(2):137–43.CrossRefGoogle Scholar
  9. 9.
    Tomita M, Wakabayashi H, Shin K, Yamauchi K, Yaeshima T, Iwatsuki K. Twenty-five years of research on bovine lactoferrin applications. Biochimie. 2009;91(1):52–7.CrossRefGoogle Scholar
  10. 10.
    Kuo Y-C, Chao I-W. Conjugation of melanotransferrin antibody on solid lipid nanoparticles for mediating brain cancer malignancy. Biotechnol Prog. 2016;32(2):480–90.CrossRefGoogle Scholar
  11. 11.
    Farooqi IS, Jebb SA, Langmack G, Lawrence E, Cheetham CH, Prentice AM, et al. Effects of recombinant leptin therapy in a child with congenital leptin deficiency. N Engl J Med. 1999;341(12):879–84.CrossRefGoogle Scholar
  12. 12.
    Jefferies WA, Brandon MR, Hunt SV, Williams AF, Gatter KC, Mason DY. Transferrin receptor on endothelium of brain capillaries. Nature. 1984;312(5990):162–3.CrossRefGoogle Scholar
  13. 13.
    Shindelman JE, Ortmeyer AE, Sussman HH. Demonstration of the transferrin receptor in human breast cancer tissue. Potential marker for identifying dividing cells. Int J Cancer. 1981;27(3):329–34.CrossRefGoogle Scholar
  14. 14.
    Walker RA, Day SJ. Transferrin receptor expression in non-malignant and malignant human breast tissue. J Pathol. 1986;148(3):217–24.CrossRefGoogle Scholar
  15. 15.
    Dixit S, Novak T, Miller K, Zhu Y, Kenney ME, Broome A-M. Transferrin receptor-targeted theranostic gold nanoparticles for photosensitizer delivery in brain tumors. Nano. 2015;7(5):1782–90.Google Scholar
  16. 16.
    Singh R, Norret M, House MJ, Galabura Y, Bradshaw M, Ho D, et al. Dose-dependent therapeutic distinction between active and passive targeting revealed using transferrin-coated PGMA nanoparticles. Small. 2016;12(3):351–9.CrossRefGoogle Scholar
  17. 17.
    Liu K, Dai L, Li C, Liu J, Wang L, Lei J. Self-assembled targeted nanoparticles based on transferrin-modified eight-arm-polyethylene glycol–dihydroartemisinin conjugate. Sci Rep. 2016;6:29461–72.CrossRefGoogle Scholar
  18. 18.
    Lee JH, Engler JA, Collawn JF, Moore BA. Receptor mediated uptake of peptides that bind the human transferrin receptor. Eur J Biochem. 2001;268(7):2004–12.CrossRefGoogle Scholar
  19. 19.
    Kim S, Choi SJ. Identification of a transferrin receptor binding peptide from a phage-displayed peptide library. J Life Sci. 2008;18(3):298–303.CrossRefGoogle Scholar
  20. 20.
    Dai XY, Xiong YL, Xu DD, Li LY, Su ZJ, Zhang QH, et al. TfR binding peptide screened by phage display technology—characterization to target cancer cells. Trop J Pharm Res. 2014;13(3):331–8.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Yuyu Tan
    • 1
  • Wenli Liu
    • 1
  • Zhi Zhu
    • 1
  • Lijun Lang
    • 1
  • Junxia Wang
    • 1
  • Mengjiao Huang
    • 1
  • Mingxia Zhang
    • 1
  • Chaoyong Yang
    • 1
  1. 1.MOE Key Laboratory of Spectrochemical Analysis and Instrumentation, Collaborative Innovation Center of Chemistry for Energy Materials, Key Laboratory for Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, College of Chemistry and Chemical EngineeringXiamen UniversityXiamenChina

Personalised recommendations