Microfluidic chip or biomolecule crystallization

Abstract

A system for crystallizing a protein and other biomolecules comprises a flow channel for conveying a solution containing the biomolecule to be crystallized and one or more substations related to the crystallization process formed along the flow channel. The system operates by flowing a biomolecule solution through a the flow channel, then performing dialysis on a micro-scale to set crystallization conditions. After dialysis, the concentration of the biomolecule solution is changed by removing water or solvent from the biomolecule solution to promote formation of crystals in a segment of the flow channel. Formed crystals may be observed and harvested from the flow channel.

1 . A method of crystallizing a biomolecule, comprising the steps of: 
 flowing a biomolecule solution containing a biomolecule to be crystallized through a biomolecule channel; and    dialyzing the biomolecule solution through a membrane in the biomolecule channel to form a crystal of the biomolecule.    
     
     
         2 . The method of  claim 1 , wherein the step of dialyzing the biomolecule solution sets effectors in the protein solution to promote crystallization of the biomolecule.  
     
     
         3 . The method of  claim 2 , wherein the effectors comprise ions exchanged through the membrane.  
     
     
         4 . The method of  claim 2 , wherein the effectors comprise the pH of the protein solution.  
     
     
         5 . The method of  claim 2 , wherein the effectors comprise the temperature of the protein solution.  
     
     
         6 . The method of  claim 2 , wherein the effectors comprise detergents that are exchanged with the protein solution through the membrane.  
     
     
         7 . The method of  claim 1 , further comprising the step of observing the crystal formation.  
     
     
         8 . The method of  claim 1 , further comprising the step of harvesting the crystal formed during observation.  
     
     
         9 . The method of  claim 1 , further comprising the step of freezing the crystal for x-ray diffraction.  
     
     
         10 . A method of crystallizing biomolecules, comprising the steps of: 
 flowing a biomolecule solution containing a biomolecule to be crystallized through a biomolecule channel; and    concentrating the biomolecule by removing solvent from the biomolecule solution at a concentration station formed along the biomolecule channel.    
     
     
         11 . The method of  claim 10 , wherein the step of concentrating comprises evaporating the solvent from the biomolecule solution through an aperture formed in a sidewall of the biomolecule channel.  
     
     
         12 . The method of  claim 10 , wherein the step of concentrating comprises filtering the biomolecule solution through a biomolecule impermeable membrane to remove the solvent from the biomolecule solution.  
     
     
         13 . The method of  claim 10 , further comprising the step of observing the crystal formation.  
     
     
         14 . The method of  claim 10 , further comprising the step of harvesting the crystal formed during observation.  
     
     
         15 . The method of  claim 10 , further comprising the step of freezing the crystal for x-ray diffraction.  
     
     
         16 . A method of crystallizing a biomolecule, comprising the steps of: 
 flowing a biomolecule solution containing a biomolecule to be crystallized through the biomolecule channel;    dialyzing the biomolecule solution through a membrane to set effectors in the biomolecule solution; and    concentrating the biomolecule by removing solvent from the biomolecule solution at an concentration station formed along the biomolecule channel, whereby a crystal of the biomolecule is formed in the biomolecule channel.    
     
     
         17 . The method of  claim 16 , further comprising the step of observing the crystal formation.  
     
     
         18 . The method of  claim 16 , further comprising the step of harvesting the crystal formed during observation.  
     
     
         19 . The method of  claim 16 , further comprising the step of freezing the crystal for x-ray diffraction.  
     
     
         20 . A system for crystallizing a biomolecule, comprising: 
 a substrate;    a microchannel formed in the substrate for conveying a biomolecule solution containing a biomolecule to be crystallized; and    a concentration substation formed in the substrate along the first microchannel for concentrating the biomolecule solution to promote crystallization of the biomolecule.    
     
     
         21 . A system for crystallizing a biomolecule, comprising: 
 a membrane for exchanging solutes between a first chamber on a first side of the membrane and a second chamber on a second side of the membrane;    a first microchannel for transmitting a biomolecule solution containing a biomolecule to be crystallized to the first chamber;    a second microchannel for delivering a buffer solution to the second chamber, wherein the biomolecule solution exchanges solvents with the buffer solution to promote crystallization, and    a first flow source in communication with the first microchannel for inducing a flow in the biomolecule solution.    
     
     
         22 . A system for crystallizing a biomolecule, comprising: 
 a substrate;    a first microchannel in the substrate for conveying a biomolecule solution containing a molecule to be crystallized;    a second microchannel intersecting the first microchannel for conveying a buffer solution for promoting crystallization of the molecule;    a recess formed in the substrate in communication with the second microchannel;    a membrane for exchanging solutes between the first microchannel and the second microchannel; and    a cap disposed above the membrane to define a crystallization chamber.    
     
     
         23 . A microfluidic chip for crystallizing a biomolecule, comprising: 
 a substrate;    an effector setting substation formed in the substrate for setting effectors in a biomolecule solution containing the biomolecule to be crystallized to promote crystallization of the biomolecule;    a concentration substation formed in the substrate for concentrating the biomolecule solution;    a detection substation formed in the substrate for detecting a crystal formed in the biomolecule solution; and    a harvesting substation for removing the crystal from the microfluidic chip.    
     
     
         24 . A system for crystallizing a biomolecule comprising: 
 a main channel through which the biomolecule solution flows;    an effector setting station formed along the main channel for selective exchange of solutes of particular size between a buffer solution and the biomolecule solution to promote crystallization of the biomolecule;    a concentration station formed along the main channel for controlled removal of solvent from the biomolecule solution;    an observation station for observing a crystal formed in the main channel; and    a crystal harvesting station for removing a formed crystal from the main channel.    
     
     
         25 . A crystallization system for crystallizing a biomolecule, comprising: 
 a substrate,    a biomolecule channel for conveying a biomolecule solution containing a biomolecule to be crystallized; and    a plurality of crystallization regions formed in series along the biomolecule channel for crystallizing the biomolecule.    
     
     
         26 . The system of  claim 25 , further comprising: 
 one or more condition re-setting substations between each crystallization region for re-setting conditions in the biomolecule channel after the biomolecule solution passes through each crystallization region.    
     
     
         27 . The system of  claim 25 , further comprising: 
 an observation station formed along the biomolecule channel for observing crystallization of the biomolecule.    
     
     
         28 . The system of  claim 25 , further comprising: 
 a harvesting station formed along the biomolecule channel for extracting a formed crystal from the channel.    
     
     
         29 . A system for crystallizing a biomolecule, comprising: 
 a channel for conveying a crystal formed of the biomolecule; and    an observation station formed along said channel for observing the crystal.    
     
     
         30 . The system of  claim 29 , further comprising: 
 a harvesting station for removing the crystal from the channel.    
     
     
         31 . The system of  claim 30 , wherein the harvesting station comprises: 
 an aperture formed in a sidewall of the channel; and    a harvesting pin configured to be inserted into said aperture for acquiring the crystal on the pin.    
     
     
         32 . The system of  claim 31 , wherein the harvesting pin comprises a first tip and a second tip spaced from the first tip to form a crystal acquisition region for holding the crystal.  
     
     
         33 . A method of handling a crystal, comprising: 
 forming a crystal in a channel;    removing the crystal from the channel using a harvesting device; and    freezing the crystal on the harvesting device.    
     
     
         34 . The method of  claim 33 , wherein the harvesting device comprises a first pin having a first tip and a second pin having a second tip spaced from the first tip to form a crystal acquisition region for holding the crystal.  
     
     
         35 . A microfluidic system for crystallizing biomolecules, comprising: 
 a biomolecule channel formed in a substrate for conveying a biomolecule solution;    a first crystallization region formed along the biomolecule channel for promoting crystallization of a biomolecule in the biomolecule solution; and    a second crystallization region formed along the biomolecule channel and in series with the first crystallization region for promoting crystallization of a biomolecule in the biomolecule solution.    
     
     
         36 . The system of  claim 35 , further comprising: 
 a dialysis station formed between the first crystallization region and the second crystallization region for dialyzing the biomolecule solution.    
     
     
         37 . A method of manufacturing a biomolecule crystallization system, comprising: 
 providing a substrate;    forming a channel in the substrate; and    providing a substation along the channel for processing a biomolecule solution in the channel.    
     
     
         38 . A collective rational drug design method, comprising: utilizing parameters derived from a family of protein crystal structures such that a drug is designed.  
     
     
         39 . In a rational drug design method wherein the improvement comprises, the use of a family of protein structures.  
     
     
         40 . A crystallized polypeptide having a integration ratio of less than 1:2500.  
     
     
         41 . A crystallized polypeptide formed under dynamic conditions.  
     
     
         42 . The crystallized polypeptide of  claim 41 , wherein said polypeptide is useful in structural determination.  
     
     
         43 . A crystallized polypeptide formed using the method of  claim 1 .  
     
     
         44 . The crystallized polypeptide of  claim 43 , wherein said crystallized polypeptide is useful in structural determination.  
     
     
         45 . The crystallized polypeptide of  claim 43 , wherein said crystallized polypeptide is formed under static conditions.  
     
     
         46 . The crystallized polypeptide of  claim 43 , wherein said crystallized polypeptide is formed under dynamic conditions.