Array for crystallizing protein, device for crystallizing protein and method of screening protein crystallization using the same

• US 20050075482A1
• Filed: 12/11/2002
• Published: 04/07/2005
• Est. Priority Date: 12/11/2001
• Status: Abandoned Application

• Current Assignee: Mitsubishi Rayon Company Limited (Mitsubishi Chemical Holdings Corporation)
• Original Assignee: Mitsubishi Rayon Company Limited (Mitsubishi Chemical Holdings Corporation)
• Inventors: Watanabe, Nobuhisa, Tanaka, Isao, Sumi, Toshinori, Nishijima, Chiharu, Nagata, Yuichirou, Takeuchi, Hiroshi, Akita, Takashi
• Application Number:
  US10/498,232
• Publication Number:
  US 20050075482A1
• Time in Patent Office: Days
• US Class Current: 530/350
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00522 in a multiple parallel arrangement
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00639 the compounds being trapped in or bound to a porous medium
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00673 Slice arrays
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00745 Inorganic compounds
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 3/06 Crystallising dishes
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07K 1/306 by crystallization
• CPC Class Codes: C30B 29/58 Macromolecular compounds
• CPC Class Codes: C30B 7/00 : Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions from molten solvents C30B9/00; by normal or gradient freezing C30B11/00; under a protective fluid C30B27/00
• CPC Class Codes: C40B 40/18 : Libraries containing only inorganic compounds or inorganic materials
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 25/147 : by cristallisation

Abstract

The present invention relates to a method of precipitating protein crystals from a protein-containing sample. The present invention also relates to a novel microarray and a novel device for screening for protein crystallization condition. Furthermore, the present invention relates to a method of conveniently and quickly screening for protein crystallization conditions using the microarray or the device having highly integrated and held crystallization conditions even with an extremely small quantity of a sample.

Claims

1. A method of crystallizing protein, which comprises the following steps of:

(a) applying a protein-containing sample to polymer gel holding protein crystallization agents; and

(b) bringing the protein into contact with the protein crystallization agents.

2. The method of crystallizing protein of claim 1, wherein the protein-containing sample contains protein, a protein solubilizing agent, and/or an additive.

3. The method of crystallizing protein of claim 1, wherein the protein crystallization agent contains a precipitating agent and/or a pH buffering agent.

4. The method of crystallizing protein of claim 1, wherein the polymer gel holding the protein crystallization agents is transparent.

5. A microarray for protein crystallization, which comprises at least 10 different types and/or different concentrations of protein crystallization agents held thereon per 1 cm² of the microarray surface.

6. The microarray of claim 5, wherein the protein crystallization agent contain a precipitating agent and/or a pH buffering agent.

7. The microarray of claim 5 or 6, wherein the protein crystallization agents are held by means of the polymer gel.

8. The microarray of claim 7, wherein the polymer gel holding the protein crystallization agents is transparent.

9. The microarray of claim 5, which comprises a plurality of hollow fibers arrayed therein holding different types and/or different concentrations of the protein crystallization agents in the hollow portions thereof.

10. The microarray of claim 9, wherein the protein crystallization agents are held by means of the polymer gel that fills the hollow portions.

11. The microarray of claim 9 or 10, wherein the protein crystallization agent contains a precipitating agent and/or a pH buffering agent.

12. A device for protein crystallization, which is provided with the following (a) and (b):

(a) a microarray for protein crystallization having protein-crystallization-agent-holding portions that comprise at least 10 different types and/or different concentrations of protein crystallization agents held thereon per 1 cm² of the microarray surface; and

(b) a plate having wells that can be filled with a protein-containing sample, and correspond to each protein-crystallization-agent-holding portion of the microarray (a) for protein crystallization.

13. The device for protein crystallization of claim 12, wherein in (a), the protein crystallization agents are held by means of polymer gel.

14. The device for protein crystallization of claim 12 or 13, wherein in (a), the protein crystallization agent contains a precipitating agent and/or a pH buffering agent.

15. The device for protein crystallization of claim 12, wherein in (a), the microarray for protein crystallization comprises a plurality of hollow fibers arrayed thereon holding different types and/or different concentrations of the protein crystallization agents in the hollow portions thereof.

16. The device for protein crystallization of claim 12, wherein in (b), the capacity of the well is 1 μ

L or less.

17. The device for protein crystallization of claim 12, wherein in (b), when superposed on the microarray (a) for protein crystallization, the superposed portion corresponding to each protein-crystallization-agent-holding portion of the array is transparent.

18. The device for protein crystallization of claim 12, wherein in (b), the plate is prepared from an optically transparent material.

19. The device for protein crystallization of claim 18, wherein the plate is made of a transparent resin or glass.

20. The device for protein crystallization of claim 12, wherein in (b), the plate has a structure in which a plate material perforated regularly is adhered to a substrate.

21. The device for protein crystallization of claim 20, wherein the substrate is made of an optically transparent material.

22. The device for protein crystallization of claim 20, wherein the substrate is prepared from a slide glass.

23. The device for protein crystallization of claim 20, wherein the plate material is made of material selected from the group consisting of metal, resin, and rubber.

24. The device for protein crystallization of claim 12, wherein the microarray (a) for protein crystallization is fixed on a support.

25. The device for protein crystallization of claim 24, wherein the support is made of an optically transparent material.

26. The device for protein crystallization of claim 24, wherein the support is prepared from a slide glass.

27. The device for protein crystallization of claim 24, wherein marking is made on the support so as to determine a position for fixing the microarray (a) for protein crystallization.

28. The device for protein crystallization of claim 24, wherein a spacer is placed between the support and the plate (b), the spacer having a space into which the microarray (a) for protein crystallization fixed on the support can be accommodated.

29. The device for protein crystallization of claim 28, wherein the spacer is made of material selected from the group consisting of metal, resin, and rubber.

30. A method of screening for protein crystallization conditions, which comprises a step of precipitating protein by bringing the protein crystallization agents into contact with a protein-containing sample on and/or in the microarray, using the microarray claim 5, or the device of claim 12.

31. The method of claim 30, wherein the protein-containing sample contains protein, a protein solubilizing agent, and/or an additive.

32. The method of claim 30, which is conducted under a plurality of temperature conditions.

33. The method of claim 30, wherein the volume of the protein-containing sample solution is 1 μ

L or less per type or concentration of the protein crystallization agent.

34. A method of screening for protein crystallization conditions, which comprises a step of precipitating protein by superposing the wells of (b) filled with the protein-containing sample onto the protein-crystallization-agent-holding portions holding the protein crystallization agents on the microarray (a), so as to bring the protein crystallization agents into contact with the protein in the device for protein crystallization of claim 12.