Kwiatkowska Eliza P, Kazimierczak Urszula, Mackiewicz Andrzej, Kowalczyk Dariusz W
Department of Cancer Diagnostics and Immunology, GreatPoland Cancer Center, Poznań, Poland.
Acta Biochim Pol. 2006;53(2):361-9. Epub 2006 May 30.
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.
我们已构建并在NS0小鼠骨髓瘤细胞中表达了与人IgG1的Fc部分融合的重组嵌合可溶性转化生长因子βII型受体（sTβRII-Fc），并分离出持续分泌非常高水平生物活性蛋白的细胞系。GS-NS0表达系统利用了强大的人巨细胞病毒立即早期启动子表达载体以及谷氨酰胺合成酶作为选择标记。重组嵌合受体能够大量产生，并通过在蛋白A柱上进行一步层析高效纯化。生化研究表明，重组sTβRII-Fc可结合天然的转化生长因子β1和转化生长因子β3亚型，并在体外中和它们的活性。
