公开(公告)号：US20130288908A1

公开(公告)日：2013-10-31

申请号：US13990654

申请日：2011-11-30

Applicant: Yasuhiro Fujino ,  Risako Fujita ,  Kouichi Wada ,  Kotomi Oda ,  Takuya Ueda ,  Yoshihiro Shimizu ,  Takashi Kanamori

Inventor： Yasuhiro Fujino ,  Risako Fujita ,  Kouichi Wada ,  Kotomi Oda ,  Takuya Ueda ,  Yoshihiro Shimizu ,  Takashi Kanamori

IPC: G01N33/68

CPC classification number: G01N33/6854 ,  C07K16/468 ,  C07K2317/31 ,  C07K2317/55 ,  C12N15/1041 ,  C12N15/1062 ,  C12N15/1075 ,  G01N33/6857

Abstract: 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.

Abstract translation: 以下的（1）或（2）的多核苷酸构建体用于进行核糖体展示，CIS显示和/或mRNA显示，以便针对感兴趣的抗原筛选Fab：（1）单核细胞系包含核糖体 - 结合序列，Fab第一链编码序列，接头肽编码序列，Fab第二链编码序列和支架编码序列，并且在其3'末端还包含与Fab保持复合物所必需的结构 其自身编码，和（2）包含Fab第一链表达顺反子和Fab第二链表达顺反子的多核苷酸构建体，其各自含有核糖体结合序列，Fab第一链编码序列或Fab第二链编码序列 和支架编码序列，所述第一Fab表达顺反子在其3'末端进一步包含核糖体失速序列，所述Fab第二链表达顺反子在其3'末端还包含结构ne 维持与Fab自身编码的复合物的塞车。

公开(公告)号：US09753040B2

公开(公告)日：2017-09-05

申请号：US13990654

申请日：2011-11-30

Applicant: Yasuhiro Fujino ,  Risako Fujita ,  Kouichi Wada ,  Kotomi Oda ,  Takuya Ueda ,  Yoshihiro Shimizu ,  Takashi Kanamori

Inventor： Yasuhiro Fujino ,  Risako Fujita ,  Kouichi Wada ,  Kotomi Oda ,  Takuya Ueda ,  Yoshihiro Shimizu ,  Takashi Kanamori

IPC: G01N33/68 ,  C12N15/10 ,  C07K16/46

CPC classification number: G01N33/6854 ,  C07K16/468 ,  C07K2317/31 ,  C07K2317/55 ,  C12N15/1041 ,  C12N15/1062 ,  C12N15/1075 ,  G01N33/6857

Abstract: 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.