Method for screening crystallization conditions in solution crystal growth

• US 20070202602A1
• Filed: 12/15/2006
• Published: 08/30/2007
• Est. Priority Date: 04/06/1999
• Status: Active Grant

• Current Assignee: Lawrence Delucas, Terry Bray
• Original Assignee: Lawrence Delucas, Terry Bray
• Inventors: DeLucas, Lawrence, Bray, Terry
• Granted Patent: US 7,700,363 B2
• Time in Patent Office: Days
• US Class Current: 436/4
• 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

1-31. -31. (canceled)

32. 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.

33. The method of claim 32 wherein said precipitate solution comprises a component selected from the group consisting of buffers, surface active agents, salts, alcohols, polyethylene glycol and mixtures thereof.

34. The method of claim 32 wherein said protein solution is buffered.

35. The method of claim 32 wherein said microarray comprises valves between protein solution microchambers and precipitate solution microchambers.

36. The method of claim 35 wherein said valves control said liquid-liquid communication between microchambers comprising protein solution and a microchambers comprising a precipitate solution.

37. The method of claim 36 wherein controlling protein crystal growth further comprises employing microchambers of varying dimensions.

38. The method of claim 32 wherein said precipitate solution and said protein solution are in fluid communication via micro-channels.

39. The method of claim 35 wherein said precipitate solution and said protein solution are in fluid communication via a channel.

40. The method of claim 36 wherein said allowing liquid-liquid communication of said precipitate solution microchambers and said protein solution microchambers comprises opening said valves.

43. The method of claim 44 wherein the platform is a microarray.

44. The method of claim 32 wherein said microarray comprises miniaturized active control elements between protein solution microchambers and precipitate solution microchambers.

45. The method of claim 44 wherein the miniaturized active control elements are selected from the group consisting of valves, pumps, and electrodes.

41. 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.

42. The method of claim 41 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.