[Fusion and expression of the gene encoding human Mn-SOD to anti-CEA single-chain antibody in Escherichia coli]

Sheng Wu Gong Cheng Xue Bao. 2000 Sep;16(5):566-9.
[Article in Chinese]

Abstract

The gene encoding human manganese-superoxide dismutase (Mn-SOD) was fused to anti-carcinoembryonic antigen single-chain antibody gene to construct the fusion gene, then was ligated into prokaryotic expression vector pET-22b(+), The fusion gene was expressed in E. coli at high level, accounting for 24% of the total bacteria soluble protein; and was characterized by SDS-PAGE and Western-blot analysis; the expression product had the CEA-binding ability in RIA, and also had the SOD activity by pyrogallol autoxidation assay. So, the Mn-SOD moiety retains substantial enzymatic activity, where the ScFv moiety can deliver the fusion protein to tumor, Mn-SOD is a potential tumor-suppressor gene, maybe the fusion protein can provide a new pathway to tumor therapy.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Carcinoembryonic Antigen / immunology*
  • Escherichia coli / genetics*
  • Humans
  • Immunoglobulin Fragments / genetics*
  • Molecular Sequence Data
  • Recombinant Fusion Proteins / biosynthesis*
  • Superoxide Dismutase / genetics*

Substances

  • Carcinoembryonic Antigen
  • Immunoglobulin Fragments
  • Recombinant Fusion Proteins
  • immunoglobulin Fv
  • Superoxide Dismutase