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

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• 作者：Ken-Ichiro  ; Ono¹ ; Masamichi  ; Kamihira  ; ;   ; ¹ ;   ; ³ ;   ; ^{kamihira@nubio.nagoya-u.ac.jp} ; Yuko  ; Kuga¹ ; Hiroyuki  ; Matsumoto¹ ; Akitsu  ; Hotta¹ ; Toshinari  ; Itoh¹ ; Ken-Ichi  ; Nishijima¹ ; Naoto  ; Nakamura² ; Haruo  ; Matsuda² ; Shinji  ; Iijima¹
• 关键词：prion protein ; scFv-Fc ; fusion protein ; chicken monoclonal antibody ; animal cell culture ; Chinese hamster ovary (CHO) ; retroviral vector ; VSV-G pseudotype
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：2003
• 出版时间：2003
• 年：2003
• 卷：95
• 期：3
• 页码：231-238
• 全文大小：1306 K

文摘

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.