Mammalian cell expression of dimeric small immune proteins (SIP)

Protein Eng. 1997 Jun;10(6):731-6. doi: 10.1093/protein/10.6.731.

Abstract

We have designed and expressed bivalent small immune proteins (SIP) based on scFv fragments connected through a short linker of four amino acids to the CH3 domain of the human immunoglobulin gamma 1 H-chain. Three different versions have been designed and expressed in mammalian cells. In one construct a cysteine residue was included in the last amino acid of the flexible 15-amino acid long linker connecting the V(L) and V(H) domains, thus creating a disulphide bond stabilized molecule. A version with a shorter (five amino acids) V(L)/V(H) linker was also produced and shown to be efficiently assembled and secreted. All three SIPs form dimers retaining their antigenic specificity in Western blotting and having a comparable functional affinity (avidity) as determined by ELISA.

MeSH terms

  • Animals
  • Blotting, Western
  • CHO Cells
  • Cricetinae
  • Dimerization
  • Enzyme-Linked Immunosorbent Assay
  • Genetic Vectors / chemical synthesis
  • Humans
  • Immunoglobulin Fragments / biosynthesis
  • Immunoglobulin Fragments / chemistry
  • Immunoglobulin Fragments / genetics*
  • Immunoglobulin Variable Region / biosynthesis
  • Immunoglobulin Variable Region / chemistry
  • Immunoglobulin Variable Region / genetics*
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Protein Biosynthesis
  • Proteins / genetics*
  • Proteins / immunology*
  • Tumor Cells, Cultured

Substances

  • Immunoglobulin Fragments
  • Immunoglobulin Variable Region
  • Proteins

Associated data

  • GENBANK/X98537
  • GENBANK/X98538