Apparatus and method for polymer synthesis using arrays

Abstract

A polymer synthesis apparatus ( 20 ) for building a polymer chain including a head assembly ( 21 ) having an array of nozzles ( 22 ) with each nozzle coupled to a reservoir ( 23 ) of liquid reagent ( 24 ), and a base assembly ( 25 ) having an array of reaction wells ( 26 ). A transport mechanism ( 27 ) aligns the reaction wells ( 26 ) and selected nozzles ( 22 ) for deposition of the liquid reagent ( 24 ) into selected reaction wells ( 26 ). A sliding seal ( 30 ) is positioned between the head assembly ( 21 ) and the base assembly ( 25 ) to form a common chamber ( 31 ) enclosing both the reaction well ( 26 ) and the nozzles ( 22 ) therein. A gas inlet ( 70 ) into the common chamber ( 31 ), upstream from the nozzles ( 22 ), and a gas outlet ( 71 ) out of the common chamber ( 31 ), downstream from the nozzles ( 22 ), sweeps the common chamber ( 31 ) of toxic fumes emitted by the reagents. Each reaction well ( 26 ) includes an orifice ( 74 ) extending into the well ( 26 ) which is of a size and dimension to form a capillary liquid seal to retain the reagent solution ( 76 ) in the well ( 26 ) for polymer chain growth therein. A pressure regulating device ( 82 ) is provided for controlling a pressure differential, between a first gas pressure exerted on the reaction well ( 26 ) and a second gas pressure exerted on an exit ( 80 ) of the orifice, such that upon the pressure differential exceeding a predetermined amount, the reagent solution ( 76 ) is expelled from the well ( 26 ) through the orifice ( 74 ). A method of synthesis of a polymer chain in a synthesis apparatus ( 20 ) is also included.

What is claimed is:  
     
         1 . A polymer synthesis apparatus for building a polymer chain by sequentially adding polymer units to a solid support in a liquid reagent comprising: 
 a base assembly including a reaction well, and having at least one orifice extending into said well;    at least one solid support disposed in said well for growing and immobilizing a polymer chain thereon;    reagent solution in said well in contact with said solid support and at least one polymer unit of said polymer chain affixed to said solid support;    retaining means positioned in said well, and formed and dimensioned to substantially prevent passage of said solid support through said orifice;    said orifice having an entrance into said well and an exit, and being of a size and dimension to form a capillary liquid seal to retain said reagent solution in said well to enable polymer chain growth therein when a pressure differential between a first gas pressure exerted on said reaction well and a second gas pressure exerted on said orifice exit is less than a predetermined amount; and    a pressure regulating device for controlling said pressure differential such that upon said pressure differential exceeding said predetermined amount, said reagent solution being expelled from said well through said orifice.    
     
     
         2 . The polymer synthesis apparatus as defined in  claim 1  wherein, 
 said retaining means comprises a filter member disposed in said well between said one orifice and said support structure.  
 
     
     
         3 . The polymer synthesis apparatus as defined in  claim 2  wherein, 
 said filter member is composed of a glass fiber frit.  
 
     
     
         4 . The polymer synthesis apparatus as defined in  claim 2  wherein, 
 said filter member and said orifice cooperate in a manner forming said capillary liquid seal.  
 
     
     
         5 . The polymer synthesis apparatus as defined in  claim 1  further including: 
 a top chamber mechanism cooperating with a top surface of said base assembly to form a top chamber enclosing said reaction well therein, said regulating device communicating with said top chamber to regulate said pressure differential.  
 
     
     
         6 . The polymer synthesis apparatus as defined in  claim 1  wherein, 
 said solid support is provided by controlled pore glass (CPG).  
 
     
     
         7 . The polymer synthesis apparatus as defined in  claim 1  wherein, 
 said polymer chain is an oligonucleotide.  
 
     
     
         8 . The polymer synthesis apparatus as defined in  claim 1  further including: 
 a bottom chamber mechanism cooperating with a bottom surface of said base assembly to form a bottom chamber enclosing said orifice therein, said regulating device communicating with said bottom chamber to regulate said pressure differential.  
 
     
     
         9 . A polymer synthesis apparatus for building a polymer chain by sequentially adding polymer units in a reagent solution, said apparatus comprising: 
 a head assembly having a plurality of nozzles mounted thereto in generally spaced-apart relation, each nozzle being coupled to a reservoir of liquid reagent for controlled delivery therethrough;    a base assembly having at least one reaction well;    a transport mechanism coupled to at least one of said head assembly and said base assembly to produce relative movement therebetween to position said reaction well and a selected one nozzle in alignment for deposition of a liquid reagent into said reaction well for synthesis of a polymer chain; and    a sliding seal positioned between said head assembly and said base assembly forms a common chamber enclosing both said reaction well and said nozzles therein.    
     
     
         10 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 each said nozzle terminates proximate a substantially planar bottom surface of said head assembly, and  
 said base assembly includes a substantially planar top surface in opposed relation to said head bottom surface.  
 
     
     
         11 . The polymer synthesis apparatus as defined in  claim 10  wherein, 
 said sliding seal is provided by a balloon seal having one end affixed to one of said head bottom surface and said base top surface, and an opposite end in sliding contact with the other of said base top surface and said head bottom surface.  
 
     
     
         12 . The polymer synthesis apparatus as defined in  claim 11  wherein, 
 said opposite end tapers radially inwardly toward an interior portion of said chamber in a manner such that upon increase in pressure in said chamber, the seal integrity between said opposite end and the other of said base top surface and said head bottom surface increases.  
 
     
     
         13 . The polymer synthesis apparatus as defined in  claim 11  wherein, 
 said seal opposite end includes a substantially stick-free coating or layer in sliding contact with the other of said base top surface and said head bottom surface.  
 
     
     
         14 . The polymer synthesis apparatus as defined in  claim 11  wherein, 
 said coating or layer comprises TEFLON®.  
 
     
     
         15 . The polymer synthesis apparatus as defined in  claim 11  wherein, 
 the other of said base top surface and said head bottom surface in sliding contact with said seal opposite end includes a substantially stick-free coating or layer therebetween.  
 
     
     
         16 . The polymer synthesis apparatus as defined in  claim 15  wherein, 
 said coating or layer comprises TEFLON®.  
 
     
     
         17 . The polymer synthesis apparatus as defined in  claim 12  wherein, 
 said seal one end is mounted to said base bottom surface, and said seal opposite end is in sliding contact with said base top surface.  
 
     
     
         18 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 said transport mechanism includes a stepped motor operably coupled to one of the head assembly and the base assembly for incremental movement thereof.  
 
     
     
         19 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 said transport mechanism is operably coupled to said base assembly for movement thereof.  
 
     
     
         20 . The polymer synthesis apparatus as defined in  claim 19  wherein, 
 said transport mechanism includes a stepped motor operably coupled to the base assembly for incremental substantially linear movement relative said nozzles.  
 
     
     
         21 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 said base assembly including a reaction well and at least one orifice extending into said well, said orifice having an entrance into said well and an exit, and being of a size and dimension to form a capillary liquid seal to retain said deposited liquid reagent in said well to enable polymer chain growth therein when a pressure differential between a first gas pressure in said chamber and a second gas pressure exerted on said orifice exit is less than a predetermined amount.  
 
     
     
         22 . The polymer synthesis apparatus as defined in  claim 21  further including: 
 a pressure regulating device for controlling said pressure differential such that upon said pressure differential exceeding said predetermined amount, said reagent solution being expelled from said well through said orifice.  
 
     
     
         23 . The polymer synthesis apparatus as defined in  claim 22  further including: 
 at least one solid support disposed in said well for growing and immobilizing said polymer chain thereon;  
 said liquid reagent deposited in said well in contact with said solid support and at least one polymer unit of said polymer chain affixed to said solid support; and  
 retaining means positioned in said well, and formed and dimensioned to substantially prevent passage of solid support through said orifice.  
 
     
     
         24 . The polymer synthesis apparatus as defined in  claim 23  wherein, 
 said retaining means comprises a filter member disposed in said well between said one orifice and said support structure.  
 
     
     
         25 . The polymer synthesis apparatus as defined in  claim 21  wherein, 
 said pressure regulating device includes an inlet into said common chamber positioned upstream from said nozzles, an outlet out of said common chamber positioned downstream from said nozzles, and a pressurized gas source coupled to said inlet for continuously streaming a gas from said gas source through said common chamber from said chamber upstream to said chamber downstream and out said outlet to sweep said chamber of toxic fumes emitted by said reagents.  
 
     
     
         26 . The polymer synthesis apparatus as defined in  claim 25  wherein, 
 said regulating device further includes a chamber valve coupled to said outlet controlling outflow of said gas from said chamber for one of raising said pressure differential above said predetermined amount and lowering said pressure differential below said predetermined amount.  
 
     
     
         27 . The polymer synthesis apparatus as defined in  claim 25  wherein, 
 said inlet and said outlet are defined by said head assembly.  
 
     
     
         28 . The polymer synthesis apparatus as defined in  claim 22  wherein, 
 said base assembly includes a top surface defining said well, and a opposite facing bottom surface, said orifice extending from said bottom surface to said top surface and into said well, and said synthesis apparatus further including: 
 a bottom chamber mechanism cooperating with said base bottom surface to form a bottom chamber enclosing said orifice therein, said regulating device communicating with said bottom chamber to form a vacuum for regulation of said pressure differential.  
 
 
     
     
         29 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 each said nozzle terminates proximate a substantially planar bottom surface of said head assembly,  
 said base assembly includes a substantially planar top surface in opposed relation to said head bottom surface, and  
 said sliding seal is provided by a balloon seal having one end affixed to one of said head bottom surface and said base top surface, and an opposite end in sliding contact with the other of said base top surface and said head bottom surface.  
 
     
     
         30 . The polymer synthesis apparatus as defined in  claim 29  wherein, 
 said opposite end tapers radially inwardly toward an interior portion of said chamber in a manner such that upon an increase in pressure in said chamber, the seal integrity between said opposite end and the other of said base top surface and said head bottom surface increases.  
 
     
     
         31 . The polymer synthesis apparatus as defined in  claim 9  wherein, 
 said reservoir, coupled to each said nozzle, comprises a plurality of independent reagent reservoirs each including a different reagent to be selectively and sequentially deposited in said well in a predetermined manner through a respective nozzle.  
 
     
     
         32 . The polymer synthesis apparatus as defined in  claim 31  wherein, 
 said transport mechanism includes a stepped motor operably coupled to said base assembly for incremental movement and alignment of said well with said respective nozzle for deposition of the selected reagent.  
 
     
     
         33 . The polymer synthesis apparatus as defined in  claim 31  wherein, 
 each nozzle communicates with said respective reservoir through a tube having a passageway communicably coupled therebetween, and  
 each tube includes a valve assembly in-line with said passageway to control passage of the reagent solution therethrough.  
 
     
     
         34 . The polymer synthesis apparatus as defined in  claim 33  further including: 
 control means operably coupled to said transport mechanism and each valve assembly for controlled sequential movement of said well relative each said nozzle and for deposition of said reagent solution therein.  
 
     
     
         35 . The polymer synthesis apparatus as defined in  claim 31  further including: 
 an inlet into said common chamber positioned upstream from said nozzles;  
 an outlet out of said common chamber positioned downstream from said nozzles; and  
 a pressurized gas source coupled to said inlet for continuously streaming a gas through said common chamber from said chamber upstream to said chamber downstream and out said outlet to sweep said chamber of toxic fumes emitted by said reagents.  
 
     
     
         36 . The polymer synthesis apparatus as defined in  claim 35  wherein, 
 said pressurize gas is an inert gas.  
 
     
     
         37 . The polymer synthesis apparatus as defined in  claim 36  wherein, 
 said inert gas is argon.  
 
     
     
         38 . The polymer synthesis apparatus as defined in  claim 35  wherein, 
 said inlet is positioned upstream of said nozzles by a sufficient distance such that said continuous stream is substantially laminar during passage thereby.  
 
     
     
         39 . The polymer synthesis apparatus as defined in  claim 38  wherein, 
 said independent reagent reservoirs are coupled to said nozzles in a manner aligning said reagents with increasing toxicity from said chamber upstream to said chamber downstream.  
 
     
     
         40 . The polymer synthesis apparatus as defined in  claim 31  wherein, 
 said base assembly includes a plurality of wells aligned in an array, and  
 said transport mechanism is formed to align any one nozzle with any one well.  
 
     
     
         41 . The polymer synthesis apparatus as defined in  claim 40  wherein, 
 said array is provided by a plurality of spaced-apart well rows and columns, and  
 said nozzles are aligned in an array having nozzle rows and columns spaced-apart by a distance substantially similar to the spacing between said spaced-apart well rows and columns for alignment therebetween such that selected wells may be simultaneously deposited with a selected reagent.  
 
     
     
         42 . The polymer synthesis apparatus as defined in  claim 41  wherein, 
 said transport mechanism includes a stepped motor operably coupled to said base assembly for linear incremental movement of said wells along a path substantially parallel with said plurality of well columns for alignment with respective nozzles for deposition of the selected reagent.  
 
     
     
         43 . The polymer synthesis apparatus as defined in  claim 41  wherein, 
 respective nozzles of each nozzle row are coupled to one reagent reservoir of said plurality of independent reagent reservoirs.  
 
     
     
         44 . The polymer synthesis apparatus as defined in  claim 41  wherein, 
 each nozzle communicates with said respective reagent reservoir through a tube forming a passageway and having one end coupled to said nozzle and an opposite end terminating in said reagent reservoir, and  
 each tube includes a valve assembly in-line with said respective passageway to control passage of the reagent solution therethrough.  
 
     
     
         45 . The polymer synthesis apparatus as defined in  claim 44  wherein, 
 each said independent reagent reservoir is pressurized to expel said liquid reagent therefrom through respective tubes.  
 
     
     
         46 . The polymer synthesis apparatus as defined in  claim 44  further including: 
 control means operably coupled to said transport mechanism and each valve assembly for controlled sequential movement of said well relative each said nozzle and for deposition of said reagent solution therein.  
 
     
     
         47 . The polymer synthesis apparatus as defined in  claim 41  wherein, 
 said base assembly includes an array of reaction wells and at least one respective orifice extending into a respective well, each orifice having an entrance into said well and an exit, and being of a size and dimension to form a capillary liquid seal to retain deposited liquid reagent in selected wells to enable polymer chain growth therein when a pressure differential between a first gas pressure in said chamber and a second gas pressure exerted each orifice exit is less than a predetermined amount.  
 
     
     
         48 . The polymer synthesis apparatus as defined in  claim 47  further including: 
 a pressure regulating device for controlling said pressure differential such that upon said pressure differential exceeding said predetermined amount, said reagent solution being expelled from said well through said orifice.  
 
     
     
         49 . The polymer synthesis apparatus as defined in  claim 48  wherein, 
 said pressure regulating device includes an inlet into said common chamber positioned upstream from said array of nozzles, an outlet out of said common chamber positioned downstream from said array of nozzles, and a pressurized gas source coupled to said inlet for continuously streaming a gas from said gas source through said common chamber from said chamber upstream to said chamber downstream and out said outlet to sweep said chamber of toxic fumes emitted by said reagents.  
 
     
     
         50 . The polymer synthesis apparatus as defined in  claim 49  wherein, 
 said regulating device further includes a chamber valve coupled to said outlet controlling outflow of said gas from said chamber for one of raising said pressure differential above said predetermined amount and lowering said pressure differential below said predetermined amount.  
 
     
     
         51 . The polymer synthesis apparatus as defined in  claim 50  wherein, 
 said inlet and said outlet are defined by said head assembly.  
 
     
     
         52 . The polymer synthesis apparatus as defined in  claim 51  wherein, 
 said inlet is provide d by an inlet slot extending substantially transversely across said common chamber such that said continuous stream is substantially laminar during passage by said array of nozzles.  
 
     
     
         53 . The polymer synthesis apparatus as defined in  claim 52  wherein, 
 said outlet is provided by an outlet slot extending substantially transversely across said common chamber.  
 
     
     
         54 . The polymer synthesis apparatus as defined in  claim 9  further including: 
 a frame assembly upon which said head assembly is pivotally mounted between a closed position, sealing said common chamber, and an open position, permitting access to said nozzles and said wells.  
 
     
     
         55 . The polymer synthesis apparatus as defined in  claim 54  wherein, 
 said base assembly is movable mounted to said frame assembly, and  
 said transport mechanism includes a stepped motor operably coupled to said base assembly for incremental movement and alignment of said well with said respective nozzle for deposition of the selected reagent.  
 
     
     
         56 . A polymer synthesis apparatus for building a polymer chain by sequentially adding polymer units in a liquid reagent, said apparatus comprising: 
 a frame assembly;    a base assembly movably mounted to said frame assembly and including an array of reaction wells aligned in a plurality of spaced-apart well rows and columns, each well having at least one respective orifice extending into said well;    a head assembly mounted to said frame assembly and having an array of nozzles aligned in a plurality of nozzle rows and columns spaced-apart by a distance substantially similar to the spacing between said well rows and columns for alignment therebetween, each nozzle being coupled to a reservoir of liquid reagent for controlled delivery therethrough;    a transport mechanism coupled to said base assembly for moving support, relative said head assembly, along a path substantially parallel to said nozzle columns to position at least one row of reaction wells in depository alignment with a selected row of nozzles for simultaneous deposition of a liquid reagent into selected reaction wells for synthesis of polymer chains;    a sliding seal positioned between said head assembly and said base assembly, and enclosing all said reaction wells and all said nozzles therein to form a common chamber;    each said well orifice being of a size and dimension to form a capillary liquid seal to retain said deposited liquid reagent in said selected wells to enable polymer chain growth therein when a pressure differential between a first gas pressure in said chamber and a second gas pressure exerted each orifice exit is less than a predetermined amount; and    a pressure regulating device for controlling said pressure differential such that upon said pressure differential exceeding said predetermined amount, said liquid reagent being expelled from respective wells through respective orifices.    
     
     
         57 . The polymer synthesis apparatus as defined in  claim 56  wherein, 
 said sliding seal is provided by a balloon seal having one end affixed to a bottom surface of said head assembly, and an opposite end in sliding contact with a top surface of said base assembly.  
 
     
     
         58 . The polymer synthesis apparatus as defined in  claim 57  wherein, 
 said transport mechanism includes a stepped motor operably coupled to said base assembly for linear incremental movement of said reactions wells along said path.  
 
     
     
         59 . The polymer synthesis apparatus as defined in  claim 58  wherein, 
 respective nozzles of each nozzle row are coupled to one reagent reservoir of said plurality of independent reagent reservoirs.  
 
     
     
         60 . The polymer synthesis apparatus as defined in  claim 59  wherein, 
 each nozzle communicates with said respective reagent reservoir through a tube forming a passageway and having one end coupled to said nozzle and an opposite end terminating in said reagent reservoir, and  
 each tube includes a valve assembly in-line with said respective passageway to control passage of the reagent solution therethrough.  
 
     
     
         61 . The polymer synthesis apparatus as defined in  claim 60  wherein, 
 said pressure regulating device includes an inlet into said common chamber positioned upstream from said array of nozzles, an outlet out of said common chamber positioned downstream from said array of nozzles, and a pressurized gas source coupled to said inlet for continuously streaming a gas from said gas source through said common chamber from said chamber upstream to said chamber downstream and out said outlet to sweep said chamber of toxic fumes emitted by said reagents.  
 
     
     
         62 . The polymer synthesis apparatus as defined in  claim 61  wherein, 
 said regulating device further includes a chamber valve coupled to said outlet controlling outflow of said gas from said chamber for one of raising said pressure differential above said predetermined amount and lowering said pressure differential below said predetermined amount.  
 
     
     
         63 . A method of synthesis for building a polymer chain by sequentially adding polymer units to at least one solid support for growing and immobilizing a polymer chain thereon in a liquid reagent, said method comprising the steps of: 
 A) depositing a liquid reagent in a reaction well in contact with at least one solid support and at least one polymer unit of said polymer chain affixed to said solid support, said well having at least one orifice extending into said well, and being of a size and dimension to form a capillary liquid seal to retain said reagent solution in said well to enable polymer chain growth on said solid support; and    B) applying a first gas pressure to said reaction well such that a pressure differential between said first gas pressure and a second gas pressure exerted on an exit of said orifice exceeds a predetermined amount necessary to overcome said capillary liquid seal and expel said reagent solution from said well through said orifice.    
     
     
         64 . The method of synthesis for building a polymer chain as described in  claim 63  further comprising the step of: 
 controlling said pressure differential through a pressure regulating device for one of raising said pressure differential above said predetermined amount and lowering said pressure differential below said predetermined amount.  
 
     
     
         65 . The method of synthesis for building a polymer chain as described in  claim 63  further comprising the step of: 
 repeating steps A and B to form at least one continuous chain of polymer units.  
 
     
     
         66 . The method of synthesis for building a polymer chain as described in  claim 63  wherein, said predetermined amount is between about {fraction (1/100)}th psi to about {fraction (1/10)}th psi.  
     
     
         67 . A method of synthesis of a polymer chain in a synthesis apparatus by sequentially adding polymer units thereto, said synthesis apparatus including a head assembly having a plurality of nozzles mounted thereto in generally spaced-apart relation, each nozzle being coupled to a reservoir of liquid reagent for controlled delivery therethrough, a base assembly having at least one reaction well, and a sliding seal positioned between said head assembly and said base assembly to permit relative movement therebetween, and enclosing both said reaction well and said nozzles therein to form a common chamber, said method comprising the steps of: 
 A) aligning said reaction well and a selected one nozzle through a transport mechanism coupled to at least one of said head assembly and said base assembly to produce relative movement therebetween;    B) depositing a liquid reagent into said well from said reagent reservoir through said one nozzle to enable synthesis of a polymer chain;    C) sweeping toxic fumes, emitted by said reagents, from said common chamber through passage of a gas from a pressurized gas source, coupled to an inlet into said common chamber and positioned upstream from said nozzles, and out of said chamber through an outlet out from said common chamber and positioned downstream from said nozzles.    
     
     
         68 . The method of synthesis a polymer chain as described in  claim 67  further comprising the step of: 
 D) expelling said deposited liquid reagent from said well through an orifice extending into said well by applying a first gas pressure to said common chamber such that a pressure differential between said first gas pressure and a second gas pressure exerted on an exit of said orifice exceeds a predetermined amount necessary to overcome a capillary liquid seal formed between said liquid reagent and orifice to retain said liquid reagent in said well.  
 
     
     
         69 . The method of synthesis a polymer chain as described in  claim 68  further comprising the step of: 
 repeating steps A-D to form at least one continuous chain of polymer units.  
 
     
     
         70 . The method of synthesis a polymer chain as described in  claim 68  wherein, 
 said expelling step is accomplished by controlling out flow of said gas through said chamber outlet to increase said pressure differential, by increasing first gas pressure, to a degree exceeding said predetermined amount.  
 
     
     
         71 . The method of synthesis a polymer chain as described in  claim 70  wherein, 
 said controlling step is accomplished by a chamber valve coupled to said chamber outlet for control of passage of said gas therethrough.