Method of manufacturing protein array or peptide array, method of identifying functional protein or functional peptide, protein array or peptide array, and functional protein or functional peptide identification kit

Patent metadata

Field	Value
Publication number	US-2016289671-A1
Application number	US-201414914373-A
Country	US
Kind code	A1
Filing date	Aug 5, 2014
Priority date	Aug 30, 2013
Publication date	Oct 6, 2016
Grant date	—

Abstract

There is provided a method of manufacturing a protein array or peptide array suitable for an efficient screening of a functional protein or functional peptide. The method of manufacturing a protein array or peptide array includes the steps of: (a) preparing a nucleic acid immobilized on a solid support and a cell-free synthesis system in a reactor, in which a reactor array includes the reactor having a specific aperture shape and a protein capture molecule or a peptide capture molecule provided on at least a portion of wall surface and bottom surface in the reactor; and (c) synthesizing a protein or peptide from the nucleic acid using the cell-free synthesis system and immobilizing the protein or peptide in the reactor.

First claim

1 . A method of manufacturing a protein array or peptide array, comprising:
(a) preparing a nucleic acid immobilized on a solid support and a cell-free synthesis system in a reactor, in which a reactor array includes the reactor having a specific aperture shape and a protein capture molecule or peptide capture molecule provided on at least a portion of wall surface and bottom surface in the reactor, and (c) synthesizing a protein or peptide from the nucleic acid using the cell-free synthesis system and immobilizing the protein or peptide in the reactor.

2 . The method of manufacturing a protein array or a peptide array according to claim 1 ,
wherein the nucleic acid immobilized on the solid support is one type of nucleic acid per solid support.

3 . The method of manufacturing a protein array or a peptide array according to claim 1 ,
wherein the solid support is a magnetic bead.

4 . The method of manufacturing a protein array or a peptide array according to claim 1 ,
wherein the protein capture molecule or the peptide capture molecule is a molecule having an affinity to an amino acid sequence existing in the protein or peptide.

5 . The method of manufacturing a protein array or a peptide array according to claim 1 ,
wherein the protein capture molecule or the peptide capture molecule is a nucleic acid linker having a protein linking moiety or a peptide linking moiety, and wherein the method further comprises transcribing an mRNA from the DNA using the cell-free synthesis system in the reactor, synthesizing a protein or peptide from the mRNA hybridized to the nucleic acid linker, and immobilizing the protein or peptide in the reactor.

6 . The method of manufacturing a protein array or a peptide array according to claim 1 , further comprising (b) sealing the reactor between the (a) and (c).

7 . A protein array or peptide array, manufactured by the method of manufacturing a protein array or a peptide array according to claim 1 .

8 . A method of identifying a functional protein or a functional peptide, comprising:
(d) performing a functional screening using the protein array or peptide array according to the claim 7 to specify a reactor.

9 . The method of identifying a functional protein or a functional peptide according to claim 8 , further comprising
(e) recovering a nucleic acid in the reactor.

10 . The method of identifying a functional protein or a functional peptide according to claim 9 ,
wherein the (e) superimposing a substrate having an affinity to a solid support with the reactor array, thereby printing a nucleic acid on the substrate.

11 . The method of identifying a functional protein or a functional peptide according to claim 9 ,
wherein the (e) recovering a nucleic acid in the reactor by light irradiation.

12 . The method of identifying a functional protein or a functional peptide according to claim 8 , further comprising:
(f) analyzing a base sequence of a nucleic acid corresponding to the reactor specified in the (d).

13 . A protein array or peptide array, comprising:
a reactor array comprising a reactor having a specific aperture shape; and a nucleic acid immobilized on a solid support disposed in the reactor and a protein or peptide encoded by the nucleic acid, wherein the reactor includes a protein capture molecule or peptide capture molecule for capturing the protein or peptide, the capture molecule being immobilized on at least a portion of wall surface and bottom surface in the reactor.

14 . A protein array or peptide array, comprising:
a reactor array comprising a reactor having a specific aperture shape; and a DNA disposed in the reactor and a protein or peptide encoded by the DNA, wherein the reactor comprises an mRNA synthesized from the DNA, and a nucleic acid linker having a protein linking moiety or a peptide linking moiety capturing the protein or peptide, immobilized on at least a portion of wall surface and bottom surface in the reactor.

15 . A functional protein or functional peptide identification kit, comprising:
the protein array or peptide array according to claim 13 ; and a substrate having an affinity to the solid support.

Assignees

• Univ Tokyo
• Nikon Corp

Inventors

• Ichiki Takanori
• Ueno Shingo
• Osawa Hisao

Classifications

• C07K17/14 Primary

  Peptides being immobilised on, or in, an inorganic carrier · CPC title

• C07K1/04

  on carriers {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C40B30/04

  by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding · CPC title

• G01N33/6842

  Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins · CPC title

• C12N15/1075 Primary

  by coupling phenotype to genotype, not provided for in other groups of this subclass · CPC title