Method for forming protein crystal

Patent metadata

Field	Value
Publication number	US-9708364-B2
Application number	US-201213474984-A
Country	US
Kind code	B2
Filing date	May 18, 2012
Priority date	May 18, 2011
Publication date	Jul 18, 2017
Grant date	Jul 18, 2017

Abstract

A method for producing a capsule for protein crystallization is provided. The method comprises adding a solution containing a protein and a gelling agent to an ionic cross-linking solution to form an ionically cross-linked gel capsule that encapsulates a solution of the protein.

First claim

The invention claimed is:

1. A method for producing a capsule for protein crystallization, comprising:
adding a solution containing a protein and a gelling agent to an ionic cross-linking and crystallization solution, to thereby form an ionically cross-linked gel capsule,
wherein the solution containing the protein and the gelling agent is added to the ionic cross-linking and crystallization solution by dropping,
wherein the ionically cross-linked gel capsule is maintained in the ionic cross-linking and crystallization solution to thereby produce a protein crystal encapsulated within a shell of the ionically cross-linked gel capsule, and
wherein the ionic cross-linking and crystallization solution comprises a buffer, a precipitant and a calcium ion, a concentration of the calcium ion in the ionic cross-linking and crystallization solution being higher than a concentration of a calcium ion in the solution containing the protein and the gelling agent.

2. A method for producing a protein crystal, comprising:
adding a solution containing a protein and a gelling agent to an ionic cross-linking solution to form an ionically cross-linked gel capsule, the solution containing the protein and the gelling agent being added to the ionic cross-linking solution by dropping, the ionic cross-linking solution comprising a calcium ion;
transferring the capsule to a crystallization solution, the crystallization solution comprising a buffer, a precipitant and a calcium ion, a concentration of the calcium ion in the crystallization solution being higher than a concentration of a calcium ion in the solution containing the protein and the gelling agent; and
maintaining the capsule in the crystallization solution to produce a protein crystal encapsulated within a shell of the ionically cross-linked gel capsule.

3. A method for producing a protein crystal, comprising:
providing a plate for protein crystallization, wherein the plate has one or more wells, and an ionic cross-linking and crystallization solution and a non-volatile oil covering the ionic cross-linking and crystallization solution are placed in each well; and
adding a solution containing a protein and a gelling agent to the non-volatile oil to thereby form and maintain an ionically cross-linked gel capsule in the ionic cross-linking and crystallization solution, the ionically cross-linked gel capsule having a protein crystal encapsulated within a shell of the ionically cross-linked gel capsule,
wherein the solution containing the protein and the gelling agent is added to the ionic cross-linking and crystallization solution by dropping, and
wherein the ionic cross-linking and crystallization solution comprises a buffer, a precipitant and a calcium ion, a concentration of the calcium ion in the ionic cross-linking and crystallization solution being higher than a concentration of a calcium ion in the solution containing the protein and the gelling agent.

4. The method of claim 1 , wherein the precipitant is at least one selected from the group consisting of isopropanol, 2-methyl-2,4-pentane diol, Jeffamine M-600, polyethylene glycol (PEG) 400, polyethylene glycol (PEG) 4000, and polyethylene glycol (PEG) 8000.

5. The method of claim 2 , wherein the precipitant is at least one selected from the group consisting of isopropanol, 2-methyl-2,4-pentane diol, Jeffamine M-600, polyethylene glycol (PEG) 400, polyethylene glycol (PEG) 4000, and polyethylene glycol (PEG) 8000.

6. A method for producing a protein crystal, comprising:
providing a plate for protein crystallization, wherein the plate has one or more wells, and an ionic cross-linking solution and a non-volatile oil covering the ionic cross-linking solution are placed in each well, the ionic cross-linking solution comprising a calcium ion,
adding a solution containing a protein and a gelling agent to the non-volatile oil by dropping, thereby forming an ionically cross-linked gel capsule in the ionic cross-linking solution,
transferring the capsule to a crystallization solution, the crystallization solution comprising a buffer, a precipitant and a calcium ion, a concentration of the calcium ion in the crystallization solution being higher than a concentration of a calcium ion in the solution containing the protein and the gelling agent, and
maintaining the capsule in the crystallization solution to produce a protein crystal encapsulated within a shell of the ionically cross-linked gel capsule.

7. The method of claim 3 , wherein the precipitant is at least one selected from the group consisting of isopropanol, 2-methyl-2,4-pentane diol, Jeffamine M-600, polyethylene glycol (PEG) 400, polyethylene glycol (PEG) 4000, and polyethylene glycol (PEG) 8000.

8. The method of claim 6 , wherein the precipitant is at least one selected from the group consisting of isopropanol, 2-methyl-2,4-pentane diol, Jeffamine M-600, polyethylene glycol (PEG) 400, polyethylene glycol (PEG) 4000, and polyethylene glycol (PEG) 8000.

Assignees

• Sugahara Michihiro
• Riken

Inventors

• Sugahara Michihiro

Classifications

• C07K1/306 Primary

  by crystallization · CPC title

• Y10T428/2991

  Coated · CPC title

• G01N23/207

  Diffractometry using detectors, e.g. using a probe in a central position and one or more displaceable detectors in circumferential positions · CPC title

• B65B5/04

  Packaging single articles · CPC title