US2002182637A1PendingUtilityA1

Method for screening microcrystallizations for crystal formation

47
Priority: Jun 18, 1999Filed: Dec 21, 2001Published: Dec 5, 2002
Est. expiryJun 18, 2019(expired)· nominal 20-yr term from priority
Inventors:Bernard D. SantarsieroRaymond C. StevensPeter G. SchultzJoseph M. JaklevicDerek T. YegianEarl CornellRobert Nordmeyer
B01J 2219/00378C40B 60/14B01L 3/0268B01J 2219/00702B01J 2219/00328C30B 29/58C30B 7/00Y10T436/2575B01L 2300/0829B01L 2400/0633B01J 2219/00313B01L 2300/04B01L 2300/10B01L 3/5085B01J 19/0046B01J 2219/00691B01J 2219/00756B01J 2219/00585G01N 2001/4027B01J 2219/00689B01L 3/50851B01L 2300/0609C40B 40/10C40B 70/00B01J 2219/00376B01L 3/5088B01L 3/021G01N 33/68Y10T436/25B01J 2219/00725C07K 2299/00B01J 2219/00547B01J 2219/00317B01L 2400/0478B01J 2219/0065B01J 2219/00412Y10T436/25625B01L 2300/021
47
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Claims

Abstract

A method is provided for performing array microcrystallizations to determine suitable crystallization conditions for a molecule, the method comprising: forming an array of microcrystallizations, each microcrystallization comprising a drop comprising a mother liquor solution whose composition varies within the array and a molecule to be crystallized, the drop having a volume of less than 1 microliter; storing the array of microcrystallizations under conditions suitable for molecule crystals to form in the drops in the array; and detecting molecule crystal formation in the drops by taking images of the drops.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method for performing array microcrystallizations to determine suitable crystallization conditions for a molecule, the method comprising: 
 forming an array of microcrystallizations, each microcrystallization comprising a drop comprising a mother liquor solution whose composition varies within the array and a molecule to be crystallized, the drop having a volume of less than 1 microliter;    storing the array of microcrystallizations under conditions suitable for molecule crystals to form in the drops in the array; and    detecting molecule crystal formation in the drops by taking images of the drops.    
     
     
         2 . The method according to  claim 1  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         3 . The method according to  claim 1  wherein taking images of the drops is performed without having to adjust a focus of an optical system taking the images.  
     
     
         4 . The method according to  claim 3  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         5 . The method according to  claim 1  wherein the molecule is a macromolecule.  
     
     
         6 . The method according to  claim 1  wherein the molecule is a protein.  
     
     
         7 . The method according to  claim 1  wherein the macromolecule has a molecular weight of at least 500 daltons.  
     
     
         8 . The method according to  claim 1  wherein the drop has a volume of less than about 750 nL.  
     
     
         9 . The method according to  claim 1  wherein the drop has a volume of less than about 500 nL.  
     
     
         10 . The method according to  claim 1  wherein the drop has a volume of less than about 250 nL.  
     
     
         11 . The method according to  claim 1  wherein the drop has a volume of between about 1 nL-750 nL.  
     
     
         12 . The method according to  claim 1  wherein the drop has a volume of between about 1 nL-500 nL.  
     
     
         13 . The method according to  claim 1  wherein the drop has a volume of between about 1 nL-250 nL.  
     
     
         14 . The method according to  claim 1  wherein each microcrystallization further includes a volume of mother liquor solution separate from the drop, the mother liquor solution contained in the volume having the same composition as the mother liquor solution contained in the drop, the volume comprising less than about 500 mL of the mother liquor solution.  
     
     
         15 . The method according to  claim 1  wherein each microcrystallization further includes a volume of mother liquor solution separate from the drop, the mother liquor solution contained in the volume having the same composition as the mother liquor solution contained in the drop, the volume comprising less than about 250 mL of the mother liquor solution.  
     
     
         16 . The method according to  claim 1  wherein the mother liquor solutions have at least 4 components which are varied within the array.  
     
     
         17 . The method according to  claim 1  wherein the mother liquor solutions have at least 5 components which are varied within the array.  
     
     
         18 . The method according to  claim 1  wherein the array includes greater than 96 microcrystallizations.  
     
     
         19 . The method according to  claim 1  wherein the array includes greater than 192 microcrystallizations.  
     
     
         20 . The method according to  claim 1  wherein forming the array of microcrystallizations includes using greater than 48 stock solutions to form the mother liquor solutions used in the array.  
     
     
         21 . The method according to  claim 1  wherein forming the array of microcrystallizations includes using greater than 96 stock solutions to form the mother liquor solutions used in the array.  
     
     
         22 . The method according to  claim 1  wherein forming the array of microcrystallizations includes using greater than 192 stock solutions to form the mother liquor solutions used in the array.  
     
     
         23 . The method according to  claim 1  wherein forming the array of microcrystallizations includes forming the drops within a volume range of less than about 25 nL.  
     
     
         24 . The method according to  claim 1  wherein forming the array of microcrystallizations includes forming the drops within a volume range of less than about 20 nL.  
     
     
         25 . The method according to  claim 1  wherein forming the array of microcrystallizations includes forming the drops within a volume range of less than about 15 nL.  
     
     
         26 . A method for performing array microcrystallizations to determine suitable crystallization conditions for a molecule, the method comprising: 
 forming an array of microcrystallizations, each microcrystallization comprising a hanging drop comprising a mother liquor solution whose composition varies within the array and a molecule to be crystallized, the drop having a volume of less than 1 microliter;    storing the array of microcrystallizations under conditions suitable for molecule crystals to form in the drops in the array; and    detecting molecule crystal formation in the drops by taking images of the drops.    
     
     
         27 . The method according to  claim 26  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         28 . The method according to  claim 26  wherein taking images of the drops is performed without having to adjust a focus of an optical system taking the images.  
     
     
         29 . The method according to  claim 28  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         30 . A method for performing array microcrystallizations to determine suitable crystallization conditions for a molecule, the method comprising: 
 forming an array of microcrystallizations, each microcrystallization comprising a sitting drop comprising a mother liquor solution whose composition varies within the array and a molecule to be crystallized, the drop having a volume of less than 1 microliter;    storing the array of microcrystallizations under conditions suitable for molecule crystals to form in the drops in the array; and    detecting molecule crystal formation in the drops by taking images of the drops.    
     
     
         31 . The method according to  claim 30  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         32 . The method according to  claim 30  wherein taking images of the drops is performed without having to adjust a focus of an optical system taking the images.  
     
     
         33 . The method according to  claim 32  wherein taking images of the drops comprises taking a single image of each drop.  
     
     
         34 . A method for performing array microcrystallizations to determine suitable crystallization conditions for a molecule, the method comprising: 
 forming an array of microcrystallizations, each microcrystallization comprising a microcrystallization volume comprising a mother liquor solution whose composition varies within the array and a molecule to be crystallized, the microcrystallization volume having a volume of less than  1  microliter;    storing the array of microcrystallizations under conditions suitable for molecule crystals to form in the microcrystallization volumes; and    detecting molecule crystal formation in the microcrystallization volumes by taking images of the microcrystallization volumes.    
     
     
         35 . The method according to  claim 34  wherein taking images of the microcrystallization volumes comprises taking a single image of each microcrystallization volume.  
     
     
         36 . The method according to  claim 34  wherein taking images of the microcrystallization volumes is performed without having to adjust a focus of an optical system taking the images.  
     
     
         37 . The method according to  claim 36  wherein taking images of the microcrystallization volumes comprises taking a single image of each microcrystallization volume.  
     
     
         38 . The method according to  claim 37  wherein the molecule is a macromolecule.  
     
     
         39 . The method according to  claim 37  wherein the molecule is a protein.  
     
     
         40 . The method according to  claim 37  wherein the macromolecule has a molecular weight of at least 500 daltons.  
     
     
         41 . The method according to  claim 37  wherein the micro crystallization volume has a volume of less than about 750 nL.  
     
     
         42 . The method according to  claim 37  wherein the microcrystalization volume has a volume of less than about 500 nL.  
     
     
         43 . The method according to  claim 37  wherein the microcrystalization volume has a volume of less than about 250 nL.  
     
     
         44 . The method according to  claim 37  wherein the microcrystalization volume has a volume of between about 1 nL-750 nL.  
     
     
         45 . The method according to  claim 37  wherein the microcrystalization volume has a volume of between about 1 nL-500 nL.  
     
     
         46 . The method according to  claim 37  wherein the microcrystalization volume has a volume of between about 1 nL-250 nL.  
     
     
         47 . The method according to  claim 37  wherein the mother liquor solutions have at least 4 components which are varied within the array.  
     
     
         48 . The method according to  claim 37  wherein the mother liquor solutions have at least 5 components which are varied within the array.  
     
     
         49 . The method according to  claim 37  wherein the array includes greater than 96 microcrystallizations.  
     
     
         50 . The method according to  claim 37  wherein the array includes greater than 192 microcrystallizations.  
     
     
         51 . The method according to  claim 37  wherein forming the array of microcrystallizations includes using greater than 48 stock solutions to form the mother liquor solutions used in the array.  
     
     
         52 . The method according to  claim 37  wherein forming the array of microcrystallizations includes using greater than 96 stock solutions to form the mother liquor solutions used in the array.  
     
     
         53 . The method according to  claim 37  wherein forming the array of microcrystallizations includes using greater than 192 stock solutions to form the mother liquor solutions used in the array.  
     
     
         54 . The method according to  claim 37  wherein forming the array of microcrystallizations includes forming the microcrystallization volumes within a volume range of less than about 25 nL.  
     
     
         55 . The method according to  claim 37  wherein forming the array of microcrystallizations includes forming the microcrystallization volumes within a volume range of less than about 20 nL.  
     
     
         56 . The method according to  claim 37  wherein forming the array of microcrystallizations includes forming the microcrystallization volumes within a volume range of less than about 15 nL.

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