Expression and characterization of human rheumatoid factor single-chain Fv

Yoshio Hashimoto, Takeshi Ikenaga, Kiyoshi Tanigawa, Tadashi Ueda, Ichiko Ezaki, Taiji Imoto

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

The variable region of heavy chain (V(H)) of human rheumatoid factor (hRF) IgM was connected with the variable region of light chain (V(L)) with the peptide-linker (GGGSGGGSGGGS) by genetic engineering method and the single-chain Fv (scFv) was expressed in E. coli. On design, scFv and scFv (tag) were planned; the latter had a detection marker at the carboxyl-terminal. These scFvs were expressed as inclusion bodies in E. coli, purified in the presence of 8 M urea by gel filtration and renatured to the active form in vitro. As a control, the Fv, non-covalently associated V(H) and V(L) fragments, was also constructed. The 3 derivatives showed almost the same binding activity to rabbit-IgG to which hRF is cross-reactive. ScFv (tag) was the most stable against urea among the 3 derivatives.

Original languageEnglish
Pages (from-to)941-945
Number of pages5
JournalBiological and Pharmaceutical Bulletin
Volume23
Issue number8
DOIs
Publication statusPublished - 2000
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

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