[Bioassay of the soluble human tumor necrosis factor receptor I recombinant plasmid expression in vitro].
OBJECTIVE
To detect the expression of recombinant plasmid PcDNA3.1-sTNFRI in vitro and evaluate the bioactivity of expressed sTNFRI.


METHODS
CHO cells were transfected with recombinant plasmid PcDNA3.1-sTNFRI by liposome. sTNFRI in cell culture supernatant was detected by ELISA and sTNFRI blockage of TNF-alpha cytotoxicity in L929 cells evaluated by MTT assay.


RESULTS
The expression of sTNFRI in transfected cell culture supernatant was higher than control groups (P < 0.001). The expressed sTNFRI could significantly neutralize TNF-alpha cytotoxicity in L929 cells.


CONCLUSIONS
These results showed that the recombinant plasmid PcDNA3.1-sTNFRI can be expressed in mammalian cells and the recombinant sTNFRI has biological function.
Xu
Chen-rong
CR
Department of Periodontology, Guangdong Provincial Stomatological Hospital, Guangzhou 510280, China.
Zhang
Jin-cai
JC
Zhao
Chuan-jiang
CJ
Zhang
Yun-hui
YH
D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D005822:Genetic Vectors / Q000187:drug effects; D006801:Humans; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D014162:Transfection
D000954:Antigens, Surface; D018076:DNA, Complementary; D047888:Receptors, Tumor Necrosis Factor, Type I; D011993:Recombinant Fusion Proteins