[Bioassay of the soluble human tumor necrosis factor receptor I recombinant plasmid expression in vitro]

Zhonghua Kou Qiang Yi Xue Za Zhi. 2004 May;39(3):185-8.
[Article in Chinese]

Abstract

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.

Publication types

  • English Abstract

MeSH terms

  • Animals
  • Antigens, Surface / biosynthesis
  • CHO Cells
  • Cloning, Molecular
  • Cricetinae
  • DNA, Complementary / biosynthesis
  • DNA, Complementary / genetics
  • Eukaryotic Cells / metabolism
  • Genetic Vectors / drug effects
  • Humans
  • Plasmids / biosynthesis
  • Plasmids / genetics
  • Receptors, Tumor Necrosis Factor, Type I / biosynthesis*
  • Receptors, Tumor Necrosis Factor, Type I / genetics
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / pharmacology
  • Transfection

Substances

  • Antigens, Surface
  • DNA, Complementary
  • Receptors, Tumor Necrosis Factor, Type I
  • Recombinant Fusion Proteins