Open AccessExpression of soluble recombinant TGF-beta type II receptor fused with the Fc portion of human IgG1 (sTbetaRII-Fc) in NS0 cells.
Author(s) -
Eliza Kwiatkowska,
Urszula Kazimierczak,
Andrzej Maćkiewicz,
Dariusz W. Kowalczyk
Publication year - 2006
Publication title -
acta biochimica polonica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.452
H-Index - 78
eISSN - 1734-154X
pISSN - 0001-527X
DOI - 10.18388/abp.2006_3350
Subject(s) - recombinant dna , microbiology and biotechnology , gene isoform , fusion protein , receptor , cell culture , affinity chromatography , biology , monoclonal antibody , chemistry , antibody , biochemistry , gene , enzyme , immunology , genetics
We have constructed and expressed recombinant chimeric soluble TGF-beta type II receptor fused with the Fc portion of human IgG1 (sTbetaRII-Fc) in NS0 mouse myeloma cells and isolated cell lines constitutively secreting very high levels of biologically active protein. The GS-NS0 expression system takes advantage of the strong human cytomegalovirus immediate early promoter expression vector and glutamine synthetase as a selectable marker. The recombinant chimeric receptor could be produced in high amounts and efficiently purified by one step chromatography on a protein A column. Biochemical studies revealed that recombinant sTbetaRII-Fc binds native TGF-beta1 and TGF-beta3 isoforms and neutralizes their activity in vitro.