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Title 

Expression and purification of recombinant human angiopoietin-2 produced in Chinese hamster ovary cells

Authors 

S J HwangH H ChoiK T KimHyo Jeong HongG Y KohG M Lee

Publisher 

Elsevier

Issue Date 

2005

Citation 

Protein Expression & Purification, vol. 39, no. 2, pp. 175-183

Keywords 

angiopoietinchinese hamster ovary cellsFLAG-tagTie2 receptorangiopoietin 2recombinant proteincell survivalCHO cellhamsterhuman

Abstract 

Angiopoietin-2 (Ang2) is a complex regulator of vascular remodeling that plays a role in both blood vessel sprouting and blood vessel regression through its receptor Tie2. Recombinant Chinese hamster ovary (rCHO) cell lines expressing a high level (20 μg/mL) of recombinant human Ang2 protein (rhAng2) with an amino-terminal FLAG-tag was constructed by transfecting the expression vectors into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification in medium containing stepwise increments in methotrexate level such as 0.02, 0.08, and 0.32 μM. The rhAng2 secreted from rCHO cells was purified at a purification yield of 53.6% from the cultured medium using an anti-FLAG M2 agarose affinity gel. SDS-PAGE and Western blot analyses showed that rCHO cells secret rhAng2 as a homodimeric glycoprotein form. Furthermore, rhAng2 binds to the Tie2 receptor and phosphorylates Tie2 in a concentration-dependent manner. Therefore, our rhAng2 could be useful for clarifying biological effect of exogenous Ang2 in the future.

ISSN 

1046-5928

Link 

http://dx.doi.org/10.1016/j.pep.2004.09.005

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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