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US 10,539,571 B2Granted
Method for preparing peptide fragments, kit for preparing peptide fragments to be used therein, and analysis method
AssigneeShimadzu Corporation
InventorsTakashi Shimada, Noriko Fukao, Chikage Aoki, Taka-Aki Sato
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Abstract
A method of proteolyzing a protein, including immobilizing a protein in at least one pore of a porous body, and contacting the protein immobilized in the pore and a protease immobilized on a solid surface such that the protease selectively accesses a site of the protein and proteolyzes the protein at the site.
Claims14
A kit for proteolyzing a protein, comprising:
- at least one porous body having a plurality of pores in which a protein is to be immobilized;
- and
- a plurality of particles having an average particle diameter larger than an average pore diameter of the pores,
- wherein the particles have a surface modified by a spacer such that a protease has been immobilized or is to be immobilized on the surface by specific binding with the spacer,
- a ligand molecule is immobilized in the pores of the porous body,
- the ligand molecule is capable of site-specifically interacting with and controlling an orientation of the protein to be immobilized, and
- the average particle diameter or the average pore diameter is sized such that each of the pores has a limit to the region accessible to the protease, and that the protein is to be immobilized with a tip portion extending into the region accessible to the protease.
4 more elements
A kit for proteolyzing a protein, comprising:
- at least one porous body having a plurality of pores in which a protein is immobilized;
- and
- a plurality of particles having an average particle diameter larger than an average pore diameter of the pores,
- wherein the particles have a surface on which a protease has been immobilized,
- a ligand molecule is immobilized in the pores of the porous body,
- the ligand molecule immobilized in the pores is capable of site-specifically interacting with the immobilized protein, thereby an orientation of the protein immobilized in the pores is controlled,
- the average particle diameter or the average pore diameter is sized such that the protease immobilized on the surface of the particles has access to the protein immobilized in the pores of the porous body,
- the pores each have a limit to the region accessible to the protease, and
- the protein immobilized in the pores has a tip portion extending into the region accessible to the protease.
6 more elements
Dependencies are inferred from the claim text.