Method for preparation of microarrays for screening of crystal growth conditions

• US 7,244,396 B2
• Filed: 10/23/2002
• Issued: 07/17/2007
• Est. Priority Date: 04/06/1999
• Status: Expired due to Fees

• Current Assignee: UAB Research Foundation (University of Alabama System)
• Original Assignee: UAB Research Foundation (University of Alabama System)
• Inventors: Chait, Arnon, DeLucas, Lawrence, Belgovskiy, Alexander, Stoops, Brad
• Primary Examiner(s): WARDEN, JILL ALICE
• Application Number:
  US10/278,771
• Publication Number:
  US 20030232967A1
• Time in Patent Office: 1,728 Days
• Field of Search: 356/36, 356/244, 356/246, 359/368, 359/372, 359/373, 382/128, 422/99, 422/100, 436/86, 436/807
• US Class Current: 422/501
• CPC Class Codes: B01L 3/06 Crystallising dishes
• CPC Class Codes: C07K 14/42 Lectins, e.g. concanavalin, phytohaemagglutinin
• CPC Class Codes: C07K 14/43 Sweetening agents, e.g. thaumatin, monellin
• CPC Class Codes: C12N 9/0016 with NAD or NADP as acceptor (1.4.1)
• CPC Class Codes: C12N 9/2442 Chitinase (3.2.1.14)
• CPC Class Codes: C12N 9/6427 Chymotrypsins (3.4.21.1; 3.4.21.2); Trypsin (3.4.21.4)
• CPC Class Codes: C12Y 302/01014 Chitinase (3.2.1.14)
• 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: G01N 21/01 Arrangements or apparatus for facilitating the optical investigation
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

A method of screening solution conditions suitable for the crystallization of macromolecules with a picogram to a microgram of protein using picoliter or nanoliter volumes is provided. A preferred method comprises preparing a plurality of recipe solutions in milliliter volumes, aspirating protein and recipe solutions, respectively, from a plurality of wells, and dispensing the protein and recipe solutions into a plurality of wells, covering the combined protein and recipe solutions with oil, and maintaining the solution wells until crystallization or precipitation occur therein.

Claims

1. A crystallization experiment preparation system, comprising:

a) dispense head provided with plurality of fluid dispenser tips;

b) a video camera and a long range microscope each carried on said dispense head;

c) a stage constructed and arranged to move the dispense head, the video camera, and the long range microscope together in the X, the Y, and the Z directions;

d) a deck area positioned with respect to the stage, said deck area having at least one fixture for receiving at least one recipe plate, at least one protein plate, at least one crystallization plate, a quality control area, and a wash system thereon.

2. A crystallization experiment preparation system, comprising:

a) a dispense head provided with plurality of fluid dispenser tips;

b) a video camera and a long range microscope each carried on said dispense head;

c) a deck area positioned with respect to the dispense head, said deck area having at least one fixture for receiving at least one recipe plate, at least one protein plate, at least one crystallization plate, a quality control area, and a wash system thereon;

d) wherein the dispense head, the video camera, and the long range microscope are held together in a stationary position with respect to a deck area; and

e) wherein the deck area is constructed and arranged to be moved with respect to the dispense head, the video camera, and the long range microscope by a stage movable in the X, the Y, and the Z, directions.

3. A crystallization experiment preparation system, comprising:

a) a dispense head provided with plurality of fluid dispenser tips;

b) a video camera and a long range microscope each carried on said dispense head;

c) a first stage on which the dispense head is positioned, said first stage being constructed and arranged to move the dispense head in the Z direction;

d) a deck area positioned with respect to the dispense head, said deck area having at least one fixture for receiving at least one recipe plate, at least one protein plate, at least one crystallization plate, a quality control area, and a wash system thereon; and

e) a second stage adapted to carry said deck area in the X and the Y directions.

4. The crystallization experiment preparation system of claim 3, said dispense head further comprising an aspiration and dispensing system.

5. The crystallization experiment preparation system of claim 4, said aspiration and dispensing system comprising a constant pressure fast solenoid.

6. The crystallization experiment preparation system of claim 4, said aspiration and dispensing system comprising a piezo device.

7. The crystallization experiment preparation system of claim 4, said aspiration and dispensing system comprising a syringe-driven fast solenoid.

8. The crystallization experiment preparation system of claim 3, further comprising a dispenser tip fluidics system, said dispenser tip fluidics system comprising a plurality of fluid dispenser tips.

9. The crystallization experiment preparation system of claim 8, wherein each dispenser tip of the tip fluidics system is individually calibrated.

10. The crystallization experiment preparation system of claim 9, wherein each said individually calibrated dispenser tips has an individual calibration curve used to dispense a measured and predetermined volume of a protein solution or a recipe solution there through.

11. The crystallization experiment preparation system of claim 3, further comprising at least one droplet array, each said at least one droplet array comprising at least one protein solution and at least one recipe solution prepared in blocks of protein and recipe solutions.

12. The crystallization experiment preparation system of claim 11, wherein the total time required to prepare each said protein solution and recipe solution block ranges from about one to about sixty seconds.

13. A method for screening protein crystal growth conditions comprising the steps of:

a) providing a crystallization experiment preparation system comprising a dispense head provided with plurality of fluid dispenser tips;

a video camera and a long range microscope each carried on said dispense head;

a first stage on which the dispense head is positioned, said first stage being constructed and arranged to move the dispense head in the Z direction;

a deck area positioned with respect to the dispense head, said deck area having at least one fixture for receiving at least one recipe well plate, at least one protein well plate, at least one crystallization well plate, a quality control area, and a wash system thereon; and

a second stage adapted to carry said deck area in the X and the Y directions;

b) dispensing a volume of protein solution containing a protein into at least one of the wells;

c) dispensing a recipe solution into at least one of the wells, wherein the recipe solutions was prepared from bulk ingredients prior to being dispensed into the wells and was stored in a plurality of containers or in a second multiwell plate;

d) subjecting the protein solution, or a combination solution formed by the combination of the protein solution with the recipe solution, to environmental conditions effective to form protein crystals; and

e) detecting protein crystal growth.