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Ligand-receptor assay for evaluation of functional activity of human recombinant VEGF and VEGFR-1 extracellular fragment

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cDNA encoding VEGF and Ig-like extracellular domains 2-4 of VEGFR-1 (sFlt-12-4) were cloned into prokaryotic expression vectors pET32a and pQE60. Recombinant proteins were purifi ed (metal affi nity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-12-4 showed that biotinylated VEGF specifi cally binds to the polystyrene-immobilized receptor extracellular fragment. Biotinylated recombinant sFlt-1 interacts with immobilized VEGF. Analysis of the interaction of immobilized recombinant VEGFR-1 and VEGF with C6 glioma cells labeled with CFDA-SE (vital fl uorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specifi c receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confi rmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection.

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Authors and Affiliations

  1. Department and Division of Medical Nanobiotechnologies, Medico-Biological Faculty, N. I. Pirogov Russian State Medical University, Moscow, Russia

    A. V. Leopol’d, V. P. Baklaushev, A. A. Korchagina, S. A. Shein, K. A. Pavlov & V. P. Chekhonin

  2. Department of Basic and Applied Neurobiology, V. P. Serbskii Sate Research Center of Social and Forensic Psychiatry, Ministry of Health Care and Social Development of the Russian Federation, Moscow, Russia

    V. P. Baklaushev, S. A. Shein, N. F. Grinenko, K. A. Pavlov, I. A. Ryabukhin & V. P. Chekhonin

Authors
  1. A. V. Leopol’d
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  2. V. P. Baklaushev
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  3. A. A. Korchagina
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  4. S. A. Shein
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  5. N. F. Grinenko
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  6. K. A. Pavlov
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  7. I. A. Ryabukhin
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  8. V. P. Chekhonin
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Corresponding author

Correspondence to A. V. Leopol’d.

Additional information

Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 152, No. 12, pp. 647–651, December, 2011

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Leopol’d, A.V., Baklaushev, V.P., Korchagina, A.A. et al. Ligand-receptor assay for evaluation of functional activity of human recombinant VEGF and VEGFR-1 extracellular fragment. Bull Exp Biol Med 152, 707–711 (2012). https://doi.org/10.1007/s10517-012-1612-0

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  • DOI: https://doi.org/10.1007/s10517-012-1612-0

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