Production of anti-prion scFv-Fc fusion proteins by recombinant animal cells

Ken Ichiro Ono, Masamichi Kamihira, Yuko Kuga, Hiroyuki Matsumoto, Akitsu Hotta, Toshinari Itoh, Ken Ichi Nishijima, Naoto Nakamura, Haruo Matsuda, Shinji Iijima

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27 Citations (Scopus)

Abstract

We constructed a replication-defective retroviral vector plasmid for the expression of a single-chain antibody fragment (scFv), derived from a chicken anti-human prion protein monoclonal antibody, fused with the Fc region of human IgG1. CHO-K1 and NS-1 cells were transformed with the viral vector pseudotyped with vesicular stomatitis virus G protein (VSV-G), and scFv-Fc producer clones were established. Among the established clones, CHO-2A9 cells produced a large amount of the product with an antibody-like dimerized structure in serum-free culture that facilitated the purification of scFv-Fc. The scFv-Fc specifically recognized the epitope sequence of prion protein in solid-phase enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. The injection test into quails revealed that the scFv became more stable in vivo by fusion with the Fc region. The scFv-Fc will be a useful tool for the detection of mammalian prion proteins.

Original languageEnglish
Pages (from-to)231-238
Number of pages8
JournalJournal of Bioscience and Bioengineering
Volume95
Issue number3
DOIs
Publication statusPublished - Jan 1 2003
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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    Ono, K. I., Kamihira, M., Kuga, Y., Matsumoto, H., Hotta, A., Itoh, T., Nishijima, K. I., Nakamura, N., Matsuda, H., & Iijima, S. (2003). Production of anti-prion scFv-Fc fusion proteins by recombinant animal cells. Journal of Bioscience and Bioengineering, 95(3), 231-238. https://doi.org/10.1263/jbb.95.231