Method for screening crystallization conditions in solution crystal growth

• US 7,700,363 B2
• Filed: 12/15/2006
• Issued: 04/20/2010
• 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: DeLucas, Lawrence J., Bray, Terry L.
• Primary Examiner(s): Gakh; Yelena G
• Assistant Examiner(s): Xu; Robert
• Application Number:
  US11/639,211
• Publication Number:
  US 20070202602A1
• Time in Patent Office: 1,222 Days
• Field of Search: None
• US Class Current: 436/86
• 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 2219/00313 the reactor vessels being formed by arrays of wells in blocks
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00479 Means for mixing reactants or products in the reaction vessels
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00693 Means for quality control
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01L 2300/041 Connecting closures to device or container
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 3/06 Crystallising dishes
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C30B 29/58 Macromolecular compounds
• CPC Class Codes: C30B 35/00 Apparatus not otherwise provided for, specially adapted for the growth, production or after-treatment of single crystals or of a homogeneous polycrystalline material with defined structure
• 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 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/08 : Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creation; Particular methods of cleavage from the liquid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 25/14 : by using distillation, extraction, sublimation, condensation, freezing, or crystallisation G01N25/02 takes precedence
• CPC Class Codes: G01N 33/00 : Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

A method of screening protein crystal growth conditions with picogram to microgram amounts of protein in picoliter or nanoliter volumes is provided. A preferred method comprises a microarray with a plurality of microchambers in the microarray. A protein solution is placed into the micro-chambers by an automated dispensing mechanism. The protein crystal growth conditions of each of the micro-chambers is adjusted so that the protein crystal growth conditions in at least two of the micro-chambers differs. Crystallization of the protein solution in the micro-chambers is effected. For example, crystallization can be effected by a precipitate solution and/or placing an oil barrier over the protein solution. Protein crystal growth in the micro-chambers is then observed.

Claims

1. A method of screening protein crystal growth conditions comprising:

dispensing a protein solution into protein solution microchambers of a microarray and separately dispensing precipitate solutions into precipitate solution microchambers of said microarray, said microchambers having a volume from about 0.001 nl to about 500 nl;

allowing liquid-liquid communication between protein solution microchambers and precipitate solution microchambers; and

observing said microchambers for protein crystal growth or protein precipitation.

2. The method of claim 1 wherein said precipitate solution comprises a component selected from the group consisting of buffers, surface active agents, salts, alcohols, polyethylene glycol and mixtures thereof.

3. The method of claim 1 wherein said protein solution is buffered.

4. The method of claim 1 wherein said microarray comprises valves between protein solution microchambers and precipitate solution microchambers.

5. The method of claim 4 wherein said valves control and liquid-liquid communication between microchambers comprising protein solution and a microchambers comprising a precipitate solution.

6. The method of claim 5 wherein controlling protein crystal growth further comprises employing microchambers of varying dimensions.

7. The method of claim 1 wherein said precipitate solution and said protein solution are in fluid communication via micro-channels.

8. The method of claim 4 wherein said precipitate solution and said protein solution are in fluid communication via a channel.

9. The method of claim 5 wherein said allowing liquid-liquid communication of said precipitate solution microchambers and said protein solution microchambers comprises opening said valves.

12. The method of claim 1 wherein said microarray comprises miniaturized active control elements between protein solution microchambers and precipitate solution microchambers.

13. The method of claim 12 wherein the platform is a microarray.

14. The method of claim 12 wherein the miniaturized active control elements are selected from the group consisting of valves, pumps, and electrodes.

10. A method of screening protein crystal growth conditions comprising:

transferring samples comprising a volume from about 0.001 nl to about 500 nl of a protein solution into microchambers in a platform;

providing a precipitate solution that is in liquid-liquid communication with the protein solution;

controlling the protein crystal growth conditions of the samples; and

observing a protein precipitate or protein crystals in the microchambers.

11. The method of claim 10 wherein controlling the protein crystal growth condition comprises employing a valve disposed in a microchannel between a microchamber comprising protein solution and a microchamber comprising precipitate solution.