NR ATBH
AU Ono,K.; Kamihira,M.; Kuga,Y.; Matsumoto,H.; Hotta,A.; Itoh,T.; Nishijima,K.; Nakamura,N.; Matsuda,H.; Iijima,S.
TI Production of anti-prion scFv-Fc fusion proteins by recombinant animal cells
QU Journal of Bioscience and Bioengineering 2003; 95(3): 231-8
PT journal article
AB 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.
AD Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
SP englisch
PO Japan