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Phage presentation and affinity selection of a deletion mutant of human lnterleukin-3


A deletion derivative of the cytokine human interleukin-3 (hIL-315–125, comprising amino acids 15–125 of the native protein) was produced as a fusion to the filamentous phage surface protein pIII. The cytokine was detected in association with phage particles by protein immunoblotting. Compared to an equivalent quantity of soluble cytokine, phage-presented hIL-315–125 exhibited reduced biological activity in a hIL-3-dependent cell proliferation assay. The reduction in activity was attributable to presence of phage particles in the assay, rather than directly owing to physical incorporation of the cytokine into the phage particle. Owing to the position of the amber codon in the phagemid vector, the phagemid-produced free hIL-315–125 species (designated hIL-315–125 ε) had 20 amino acids appended to its C-terminus; hIL-315–125 ε did not exhibit reduced bioactivity. hIL-315–125-presenting phage were affinity-selected with either a hIL-3-reactive polyclonal antibody or with cells expressing the heterodimeric hIL-3 receptor. These data are consistent with the use of phage-display technology for the affinity selection of hIL-3 variants with modified biological properties.

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amino acid




acute myelogenous leukemia cell




borate buffered saline


baby ham-ster kidney cell


base pair(s)


bovine serum albumin


bovine somatotropin (growth hormone)


3-(cyclohexylamino)-1-propane-sulphionic acid




counts per minute


Dulbecco’s modified Eagle’s medium


Dulbecco’s phosphate buffered saline


enzyme-linked immunosorbent assay


fetal bovine serum


human ciliary neurotrophic factor


human granulocyte colony-stimulating factor


human growth hormone


human granulocyte-macrophage colony-stimulating factor


human interleukin-3


receptor for human interleukin-3


human interleukin-5




tail protein of filamentous bacteriophage


DNA polymerase chain reaction


polyethylene glycol


sodium dodecyl sulfate


titerable unit of phage




  1. 1.

    Metcalf, D. (1993),Blood 82, 3515–3523.

    CAS  Google Scholar 

  2. 2.

    Biesma, B., Willemse, P. H. B., Mulder, N. H., Sleijfer, D. Th., Gietema, J. A., Mull, R., Limburg, P. C., Bouma, J., Vellenga, E., and de Vries, E. G. E. (1992).Blood 80, 1141–1148.

    CAS  Google Scholar 

  3. 3.

    Gianni, A. M., Siena, S., Bregni, M., Di Nicola, M., Peccatori, F., Magni, M., Ravagnani, F., Sklenar, L., and Banadonna, G. (1993).Annals Oncol. 4, 759–766.

    CAS  Google Scholar 

  4. 4.

    Zimmerman, T. M., Williams, S. F., Bender, J. G., Lee, W. J., Blake, M., Carreon, J., Swinney, P., Smith, S. J., Schilling, M., Oldham, F., Van Epps, D.,and, Williams, D.E. (1995),J. Hematotherapy 4, 527–529.

    CAS  Google Scholar 

  5. 5.

    Purdy, M. H., Hogan, C. J., Hami, L., McNeice, I., Franklin, W., Jones, R. B., Bearman, S. I., Berenson, R. J., Cagnoni, P. J., Heimfeld, S., and Shpall, E. J. (1995),J. Hematotherapy 4, 515–525.

    CAS  Google Scholar 

  6. 6.

    Miyajima, A., Mui, A. L.-F., Ogorochi T., and Kazuhiro, S. (1993).Blood 82, 1960–1974.

    CAS  Google Scholar 

  7. 7.

    Gram, H., Strittmatter, U., Lorenz, M., Glueck, D., and Zenke, G. (1993),J. Immunol. Meth. 161, 169–176.

    Article  CAS  Google Scholar 

  8. 8.

    Clackson, T. and Wells, J. A. (1994),Tibtech 12, 173–184.

    CAS  Google Scholar 

  9. 9.

    Winter, J. (1994),Drug Devel. Res. 33, 71–89.

    Article  CAS  Google Scholar 

  10. 10.

    Thomas, J. W., Baum, C. M., Hood, W. F., Klein, B., Monahan, J. B., Paik, K., Staten, N., Abrams, M., and McKearn, J. P. (1995).Proc. Natl. Acad. Sci. 92, 3779–3783.

    Article  CAS  Google Scholar 

  11. 11.

    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

  12. 12.

    Olins, P. O., Bauer, S. C., Bradford-Goldberg, S., Sterbenz, K., Polazzi, J. O., Caparon, M. H., Klein, B. K., Easton, A. M., Paik, K., Klover, J. A., Thiele, B. R., and McKearn, J. P. (1995),J. Biol. Chem. 270, 23,754–23,760.

    CAS  Google Scholar 

  13. 13.

    Parmley, S. F. and Smith, G. P. (1988),Gene 73, 305–318.

    Article  CAS  Google Scholar 

  14. 14.

    Boeke, J. D., Russel, M., and Model, P. (1980).J. Mol. Biol. 144, 103–116.

    Article  CAS  Google Scholar 

  15. 15.

    Laemmli, U. K. (1970),Nature 227, 680–685.

    Article  CAS  Google Scholar 

  16. 16.

    Bachmann, B. J. (1987), in Escherichia coliand Salmonella typhimuriumCellular and Molecular Biology, Neidhardt, F. C., Ingraham, J. L., Low, K. B., Magasanik, B., Schaechter, M., and Umbarger, H. E., eds., American Society for Microbiology, Washington, D. C., pp. 1190–1219.

    Google Scholar 

  17. 17.

    Wood, D. C., Salsgiver, W. J., Kasser, T. R., Lange, G. W., Rowold, E., Violand, B. N., Johnson, A., Leimgruber, R. M., Parr, G. R., Siegel, N. R., Kimack, N. M., Smith, C., Zobel, J. F., Ganguli, S. M., Garbow, J. R., Bild, G., and Krivi, G. G. (1989),J. Biol. Chem. 264, 14,741–14,747.

    CAS  Google Scholar 

  18. 18.

    Hippenmeyer, P. J. and Pegg, L. E. (1995),Curr. Opin. Biotech. 6, 548–552.

    Article  CAS  Google Scholar 

  19. 19.

    Vieira, J. and Messing, J. (1987),Meth. Enzymol. 153, 3–11.

    CAS  Article  Google Scholar 

  20. 20.

    Bass, S., Greene, R., and Wells, J. A. (1990),Proteins: Structure, Function and Genetics 8, 309–314.

    Article  CAS  Google Scholar 

  21. 21.

    Saggio, I., Gloaguen, L. and Laufer, R. (1995),Gene 152, 35–39.

    Article  CAS  Google Scholar 

  22. 22.

    Stent, G. S. and Calendar, R. (1978),Molecular Genetics, 2nd ed., Freeman, San Francisco.

    Google Scholar 

  23. 23.

    Roberts, D., Guegler, K., and Winter, J. (1993),Gene 128, 67–69.

    Article  CAS  Google Scholar 

  24. 24.

    Doorbar, J. and Winter, G. (1994),J. Mol. Biol. 244, 361–369.

    Article  CAS  Google Scholar 

  25. 25.

    Barry, M. A., Dower, W. J., and Johnston, S. A. (1996),Nature Medicine 2, 299–305.

    Article  CAS  Google Scholar 

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Correspondence to Stephen C. Lee.

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Merlin, S., Rowold, E., Abegg, A. et al. Phage presentation and affinity selection of a deletion mutant of human lnterleukin-3. Appl Biochem Biotechnol 67, 199–214 (1997).

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Index Entries

  • Phage display
  • cytokine
  • receptor
  • growth factor
  • proliferation
  • biopanning