fulltext.study @t Gmail

Transient expression of osteopontin in HEK 293 cells in serum-free culture

Paper ID Volume ID Publish Year Pages File Format Full-Text
17979 42708 2007 8 PDF Available
Title
Transient expression of osteopontin in HEK 293 cells in serum-free culture
Abstract

As a highly phosphorylated glycoprotein, OPN has important implications in the pathogenesis of many types of cancers and autoimmune diseases. With the green fluorescent protein (GFP) as a reporter, the parameters to transfect HEK 293-F cells were optimized. These included the concentrations of plasmid DNA and linear 25 kDa PEI, the reaction time for DNA/PEI complex formation and the time to transfect the cells. When the cloned full-length human osteopontin (OPN-a) gene was transiently expressed, a limited growth of cells and excessive accumulations of lactate and ammonium were observed in shaker flask culture. However, when compared to 27 μg/mL in shaker flask culture, the yield of OPN-a reached 49 μg/mL in a 5 L bioreactor as a result of the doubled maximal viable cell density in the presence of 1 g/L protein hydrolysates in the post-transfection growth medium. The purified OPN-a was proven to induce the cell adhesion after chromatographic purification.

Keywords
HEK 293; Osteopontin; Polyethylenimine; Purification; Transient expression
First Page Preview
Transient expression of osteopontin in HEK 293 cells in serum-free culture
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
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 41, Issues 1–2, 2 July 2007, Pages 133–140
Authors
, , , , , , , ,
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