title: Production of anti-prion scFv-Fc fusion proteins by recombinant animal cells.
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.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Ono
authors.forename: Masamichi
authors.initials: M
authors.lastname: Kamihira
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kuga
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Matsumoto
authors.forename: Akitsu
authors.initials: A
authors.lastname: Hotta
authors.forename: Toshinari
authors.initials: T
authors.lastname: Itoh
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Nishijima
authors.forename: Naoto
authors.initials: N
authors.lastname: Nakamura
authors.forename: Haruo
authors.initials: H
authors.lastname: Matsuda
authors.forename: Shinji
authors.initials: S
authors.lastname: Iijima