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

A. V. Leopol'D; V. P. Baklaushev; A. A. Korchagina; S. A. Shein; N. F. Grinenko; K. A. Pavlov; I. A. Ryabukhin; V. P. Chekhonin

doi:10.1007/s10517-012-1612-0

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

A. V. Leopol'D
, V. P. Baklaushev
, A. A. Korchagina
, S. A. Shein
, N. F. Grinenko
, K. A. Pavlov
, I. A. Ryabukhin
, V. P. Chekhonin

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

cDNA encoding VEGF and Ig-like extracellular domains 2-4 of VEGFR-1 (sFlt-1_2-4) were cloned into prokaryotic expression vectors pET32a and pQE60. Recombinant proteins were purified (metal affinity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-1_2-4 showed that biotinylated VEGF specifically 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 fluorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specific receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confirmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection.

Original language	English (US)
Pages (from-to)	707-711
Number of pages	5
Journal	Bulletin of Experimental Biology and Medicine
Volume	152
Issue number	6
DOIs	https://doi.org/10.1007/s10517-012-1612-0
State	Published - Apr 2012
Externally published	Yes

Keywords

Ligand-receptor assay
Vascular endothelial growth factor
Vascular endothelial growth factor receptor

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1007/s10517-012-1612-0

Cite this

Leopol'D, A. V., Baklaushev, V. P., Korchagina, A. A., Shein, S. A., Grinenko, N. F., Pavlov, K. A., Ryabukhin, I. A., & Chekhonin, V. P. (2012). Ligand-receptor assay for evaluation of functional activity of human recombinant VEGF and VEGFR-1 extracellular fragment. Bulletin of Experimental Biology and Medicine, 152(6), 707-711. https://doi.org/10.1007/s10517-012-1612-0

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

abstract = "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 purified (metal affinity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-12-4 showed that biotinylated VEGF specifically 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 fluorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specific receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confirmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection.",

keywords = "Ligand-receptor assay, Vascular endothelial growth factor, Vascular endothelial growth factor receptor",

author = "Leopol'D, \{A. V.\} and Baklaushev, \{V. P.\} and Korchagina, \{A. A.\} and Shein, \{S. A.\} and Grinenko, \{N. F.\} and Pavlov, \{K. A.\} and Ryabukhin, \{I. A.\} and Chekhonin, \{V. P.\}",

year = "2012",

month = apr,

doi = "10.1007/s10517-012-1612-0",

language = "English (US)",

volume = "152",

pages = "707--711",

journal = "Bulletin of Experimental Biology and Medicine",

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T1 - Ligand-receptor assay for evaluation of functional activity of human recombinant VEGF and VEGFR-1 extracellular fragment

AU - Leopol'D, A. V.

AU - Baklaushev, V. P.

AU - Korchagina, A. A.

AU - Shein, S. A.

AU - Grinenko, N. F.

AU - Pavlov, K. A.

AU - Ryabukhin, I. A.

AU - Chekhonin, V. P.

PY - 2012/4

Y1 - 2012/4

N2 - 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 purified (metal affinity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-12-4 showed that biotinylated VEGF specifically 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 fluorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specific receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confirmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection.

AB - 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 purified (metal affinity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-12-4 showed that biotinylated VEGF specifically 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 fluorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specific receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confirmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection.

KW - Ligand-receptor assay

KW - Vascular endothelial growth factor

KW - Vascular endothelial growth factor receptor

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DO - 10.1007/s10517-012-1612-0

M3 - Article

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AN - SCOPUS:85027933690

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SP - 707

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JO - Bulletin of Experimental Biology and Medicine

JF - Bulletin of Experimental Biology and Medicine

IS - 6

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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