POLYNUCLEOTIDE CONSTRUCT CAPABLE OF PRESENTING FAB IN ACELLULAR TRANSLATION SYSTEM, AND METHOD FOR MANUFACTURING AND SCREENING FAB USING SAME

摘要

The polynucleotide construct of (1) or (2) below is used to perform ribosome display, CIS display and/or mRNA display in order to screen a Fab against an antigen of interest: (1) a polynucleotide construct which monocistronically comprises a ribosome-binding sequence, Fab first chain-coding sequence, linker peptide-coding sequence, Fab second chain-coding sequence and scaffold-coding sequence in this order, and further comprises at its 3'-end a structure necessary for maintaining a complex with the Fab encoded by itself; and (2) a polynucleotide construct which comprises a Fab first chain-expressing cistron and a Fab second chain-expressing cistron each containing a ribosome-binding sequence, a Fab first chain-coding sequence or Fab second chain-coding sequence, and a scaffold-coding sequence in this order, the first Fab-expressing cistron further comprising at its 3'-end a ribosome stall sequence, said Fab second chain-expressing cistron further comprising at its 3'-end a structure necessary for maintaining a complex with the Fab encoded by itself