High levels of biologically active vascular endothelial growth factor (VEGF) are produced by the baculovirus expression system.
Vascular endothelial growth factor (VEGF) is a recently discovered mitogen for endothelial cells in vitro, and a potent angiogenesis promoting factor in vivo. VEGF is secreted from producing cells as a homodimer, binds to specific receptors on the cell surface of endothelial cells, and is produced in four forms as a result of alternative splicing. We have expressed the cDNA encoding the 165 amino-acid long isoform of VEGF in insect cells using the baculovirus based expression vector. We show that infected insect cells secrete large amounts of VEGF. Antibodies directed against a synthetic peptide prepared from human VEGF identify the secreted factor. The baculovirus derived VEGF expressed in insect cells (inVEGF) binds directly to the VEGF receptors inVEGF competes with pure mammalian cells derived [125I]-VEGF for binding to the VEGF receptors that are present on the cell surface of endothelial cells. Furthermore, inVEGF is biologically active and induces the proliferation of human umbilical vein derived endothelial cells.
Cohen
T
T
Department of Food Engineering and Biotechnology, Israel Institute of Technology, Haifa.
Gitay-Goren
H
H
Neufeld
G
G
Levi
B Z
BZ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D003001:Cloning, Molecular; D016228:Endothelial Growth Factors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004730:Endothelium, Vascular; D006801:Humans; D008222:Lymphokines / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D009036:Moths; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011975:Receptors, Mitogen / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D008222:Lymphokines; D011975:Receptors, Mitogen; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D011505:Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor