Expression of soluble recombinant TGF-beta type II receptor fused with the Fc portion of human IgG1 (sTbetaRII-Fc) in NS0 cells.
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.
Kwiatkowska
Eliza P
EP
Department of Cancer Diagnostics and Immunology, GreatPoland Cancer Center, Poznań, Poland.
Kazimierczak
Urszula
U
Mackiewicz
Andrzej
A
Kowalczyk
Dariusz W
DW
D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D007074:Immunoglobulin G / Q000235:genetics*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
D018076:DNA, Complementary; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D018125:Receptors, Transforming Growth Factor beta; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D053773:Transforming Growth Factor beta1; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II