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