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Overexpression of biologically active VEGF121 fusion proteins in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
25282 43564 2007 10 PDF Available
Title
Overexpression of biologically active VEGF121 fusion proteins in Escherichia coli
Abstract

Vascular endothelial growth factor-A (VEGF) exists as five different isoforms, which exert their growth stimulatory effects through interaction with the FLK and KDR receptors. The VEGF121 isoform has been employed as a highly selective carrier of therapeutic agents to target tumor endothelial cells resulting in inhibition of tumor growth and metastasis. VEGF121 and VEGF121/rGel fusion toxin containing hexa-histidine tags were expressed in Escherichia coli AD494 (DE3) pLysS. Media containing glycerol as a primary carbon source increased the specific expression levels of soluble VEGF121 and VEGF121/rGel (mg/L/OD10) by more than two-fold over LB media when grown in a batchtype cultivation in a bioreactor. High cell densities over OD 40 were achieved using a fed-batch method and employing feeding medium containing glycerol and yeast extract. The overall production of the target proteins was improved 18-fold for VEGF121 (59.2 mg/L) and 27-fold for VEGF121/rGel (42.5 mg/L), respectively, compared to the conventional flask cultivation method (3.3 and 1.6 mg/L for VEGF121 and VEGF121/rGel, respectively). The purified VEGF121 and VEGF121/rGel fusion proteins were biologically active as assessed by phosphorylation of KDR receptors and cytotoxicity against KDR expressing cells.

Keywords
Vascular endothelial growth factor (VEGF); VEGF121/rGel; Fed-batch cultivation; E. coli AD494 (DE3) pLysS
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Overexpression of biologically active VEGF121 fusion proteins in Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 128, Issue 3, 20 February 2007, Pages 638–647
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us