Method for crystallizing protein

Patent metadata

Field	Value
Publication number	US-10612155-B2
Application number	US-201414914755-A
Country	US
Kind code	B2
Filing date	Aug 27, 2014
Priority date	Aug 30, 2013
Publication date	Apr 7, 2020
Grant date	Apr 7, 2020

Abstract

Provided are a protein crystal device and method for crystallizing protein capable of generating protein crystal without imparting a heat effect, a protein crystal-cutting device and method for cutting protein crystal capable of cutting protein crystal without imparting a heat effect on protein crystal, and bubble-jetting member and protein-adsorbing-bubble-jetting member used in said device. A bubble-jetting member is used in a protein crystal device to jet bubbles into a protein solution to thereby allow protein crystals to be obtained, the bubble-jetting member comprising: a core formed of a conductive material; a shell part formed of an insulating material, including an extended section extending from the tip of the core, and in which at least a portion closely adheres to the core to cover the core; and a gap having a bubble-jetting port, the gap being formed between the extended section and the tip of the core.

First claim

What is claimed is:

1. A method for crystallizing protein using a protein crystal device,
the protein crystal device comprising:
a bubble-jetting member, a counter electrode, and an electricity output means,
the bubble-jetting member comprising:
a core formed of a conductive material;
a shell part formed of an insulating material, the shell part including an extended section extending from a tip of the core, and at least a portion closely adhering to the core to cover the core; and
a gap having a bubble-jetting port, the gap being formed between the extended section and the tip of the core; and

the method for crystallizing protein comprising:
a step for immersing at least the bubble-jetting port of the bubble-jetting member of the protein crystal device in a protein solution and allowing the counter electrode to make contact with the protein solution; and
a step for outputting electricity to a pair of electrodes composed of the core of the bubble-jetting member and the counter electrode, and causing bubbles to be jetted from the bubble-jetting port into the protein solution;
wherein the method for crystallizing protein is done in vitro.

2. The method for crystallizing protein using the protein crystal device of claim 1 , wherein the protein solution comprises a precipitant.

3. The method for crystallizing protein using the protein crystal device of claim 2 , wherein the precipitant is selected from PEG and Tris.

4. A method for crystallizing protein using a protein crystal device,
the protein crystal device comprising:
a protein-adsorbing-bubble-jetting member, a counter electrode, and an electricity output means,
the protein-adsorbing-bubble-jetting member comprising:
a core formed of a conductive material;
a shell part formed of an insulating material, the shell part including an extended section extending from a tip of the core, and at least a portion closely adhering to the core to cover the core;
a gap having a bubble-jetting port, the gap being formed between the extended section and the tip of the core; and
an outside shell part on the outside of the shell part;

wherein
the outside shell part has an axis coaxial with the center axis of the shell part,
the outside shell part is formed at a position away from the shell part so that a space is left therebetween, and
the outside shell part has a protein-adsorbing-bubble-jetting port,

the method for crystallizing protein comprising:
a step for introducing a protein solution into the space between the outside shell part and the shell part of the protein-adsorbing-bubble-jetting member of the protein crystal device and/or the space formed by the outside shell part and a tip of the extended section of the shell part;
a step for immersing at least the protein-adsorbing-bubble-jetting port of the protein-adsorbing-bubble-jetting member in a protein solution, buffer solution, or precipitant-containing solution, and forming a state in which the core of the protein-adsorbing-bubble-jetting member and the counter electrode can be energized; and
a step for outputting electricity to a pair of electrodes constituted by the counter electrode and the core of the protein-adsorbing-bubble-jetting member, and jetting bubbles, in which protein and/or a protein solution has been adsorbed onto the boundary, from the protein-adsorbing-bubble-jetting port into the protein solution, buffer solution, or precipitant-containing solution.

5. The method for crystallizing protein using the protein crystal device of claim 4 , wherein the protein solution is introduced into the space by a liquid feed pump, or introduced by capillary action through immersion of the protein-adsorbing-bubble-jetting port into the protein solution.

Assignees

• Japan Science & Tech Agency

Inventors

• Yamanishi Yoko
• Arai Fumihito
• KURIKI Hiroki

Classifications

• C30B30/02

  using electric fields, e.g. electrolysis · CPC title

• C12N9/2462

  Lysozyme (3.2.1.17) · CPC title

• C30B29/58

  Macromolecular compounds · CPC title

• C12N9/0016

  with NAD or NADP as acceptor (1.4.1) · CPC title

• C30B7/12 Primary

  by electrolysis · CPC title