USRE37941EExpiredUtility

Capillary electrophoresis using replaceable gels

75
Assignee: BECKMAN COULTER INCPriority: Jan 29, 1991Filed: Aug 5, 1999Granted: Dec 31, 2002
Est. expiryJan 29, 2011(expired)· nominal 20-yr term from priority
Inventors:Andras Guttman
G01N 27/44704G01N 27/44747
75
PatentIndex Score
29
Cited by
21
References
46
Claims

Abstract

The filling of an internally coated capillary with a gel in its polymerized state without damaging the gel. The coating prevents bonding of the gel to the inside of the capillary, The gel comprises up to 6% acrylamide and 0-5% crosslinker. The gel can be advantageously and conveniently used in automated electrophoresis systems for automatic replacement of spent gel.

Claims

exact text as granted — not AI-modified
I claim:  
     
       1. An apparatus for separating a sample into its monomer species comprising: 
       a capillary;  
       a supply of polymerized gel disposed in communication with one end of the capillary;  
       means for filling the capillary with the polymerized gel;  
       means for introducing the sample into the filled capillary; and  
       means for performing electrophoresis on the sample to separate into its molecular species.  
     
     
       2. A system as in  claim 1  further comprising  An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species; and 
       
       removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel.  
     
     
       3. A system as in  claim 1 An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel; and  
       
       means for automatic control of operation of the system  apparatus.  
     
     
       4. A system as in  claim 1 An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel; and  
       
       
         means for automatic control of operation of the apparatus; 
       
       wherein the polymerized gel and sample are contained in receptacles; and  
       wherein the system  apparatus further comprises a carousel for supporting the receptacles and means for turning the carousel to position the carousel to position the  place a respective receptacle in flow communication with the capillary.  
     
     
       5. A system as in  claim 4 An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel; and  
       
       
         means for automatic control of operation of the apparatus,  
       
         wherein the polymerized gel and sample are contained in receptacles and the apparatus further comprises a carousel for supporting the receptacles and means for turning the carousel to position the carousel to a respective receptacle in flow communication with the capillary, and  wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing.  
     
     
       6. A system as in  claim 5  further comprising means for automatic control of operation of the system. 
     
     
       7. A system as in  claim 6  wherein  An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel; and  
       
       
         means for automatic control of operation of the apparatus,  
       
       
         wherein the polymerized gel and sample are contained in receptacles and the apparatus further comprises a carousel for supporting the receptacles and means for turning the carousel to position the carousel to place a respective receptacle in flow communication with the capillary, and  
       
         wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and wherein  the means for automatic control of the apparatus is programmed to displace the spent gel originally in the capillary by filling with polymerized gel from the receptacle into the capillary.  
     
     
       8. A system as in  claim 7 An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel;  
       
       
         means for automatic control of an operating system, and  
       
       
         means for analyzing electrophoretically separated components of the sample,  
       
       
         wherein the polymerized gel and sample are contained in receptacles and the apparatus further comprises a carousel for supporting the receptacles and means for turning the carousel to position the carousel to place a respective receptacle in flow communication with the capillary, and  
       
         wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and wherein the means for automatic control is programmed to displace the spent gel originally in the capillary by filling with a polymerized gel from the receptacle into the capillary, and the apparatus  further comprising means for analyzing electrophoretically separated components of the sample.  
     
     
       9. A system as in  claim 8 An apparatus for separating a sample into its molecular species comprising: 
       
         a capillary and a cartridge for supporting the capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing the sample into the filled capillary;  
       
       
         means for performing electrophoresis on the sample to separate the sample into its molecular species;  
       
       
         removing means for removing a spent gel from the capillary so that it can be refilled with polymerized gel;  
       
       
         means for automatic control of an operating system, and  
       
       
         means for analyzing electrophoretically separated components of the sample,  
       
       
         wherein the polymerized gel and sample are contained in receptacles and the system further comprises a carousel for supporting the receptacles and means for turning the carousel to position a respective receptacle in flow communication with the capillary, and  
       
       
         the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and  
       
         wherein the means for automatic control is programmed to displace the spent gel originally in the capillary by filling with polymerized gel from the receptacle into the capillary, and the apparatus  further comprising  comprises a cartridge for supporting the capillary.  
     
     
       10. A system  An apparatus as in claim  1   2 wherein the capillary has inner walls coated with a material that prevents bonding of the polymerized gel to the walls. 
     
     
       11. A system  An apparatus as in  claim 10  wherein the polymerized gel has a composition comprising up to 6% acrylamide and 0-5% crosslinker . 
     
     
       12. A gel-containing capillary column useful for electrophoresis comprising: 
       a capillary having a bore coated with a material having a property which prevents bonding of gel to the material; and  
       a polymeric gel filling said bore, wherein the gel has a composition that allows it to be forced to fill the bore substantially without damage to gel structure which might otherwise significantly affect the electrophoretic performance of the gel.  
     
     
       13. A capillary column as in  claim 1  wherein the gel comprises: 
       up to 6% of acrylamide; and  
       0-5% of crosslinker.  
     
     
       14. A capillary column as in  claim 13  wherein the crosslinker comprises methylenebisacrylamide. 
     
     
       15. A capillary column as in  claim 14  wherein the coated material contains either phenyl groups and methyl groups or cyanopropyl groups. 
     
     
       16. A capillary electrophoresis system comprising: 
       a capillary;  
       a supply of polymerized gel disposed in communication with one end of the capillary;  
       a supply of sample;  
       means for filing the capillary with the gel and introducing a sample into the capillary; and  
       means for performing electrophoresis.  
     
     
       17. A system as in  claim 16  further comprising  A capillary electrophoresis system comprising: 
       
         a capillary;  
       
       
         a supply of polymerized gel disposed in communication with one end of the capillary;  
       
       
         a supply of sample;  
       
       
         means for filling the capillary with the polymerized gel;  
       
       
         means for introducing a sample into the capillary;  
       
       
         means for performing electrophoresis, and  
       
         removing  means for removing the gel from the capillary so that it can be refilled with polymerized gel.  
     
     
       18. A system as in  claim 17  further comprising means for automatic control of the operation of the system  apparatus. 
     
     
       19. A system as in claim  26   17  further comprising receptacles wherein the gel and sample are contained in receptacles and wherein the system further comprises a carousel  carrier for supporting the receptacles and means for turning the carousel  carrier to position the carousel to position the  a respective receptacle in flow communications with the capillary. 
     
     
       20. A system as in  claim 19  wherein the means for for  filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing. 
     
     
       21. A system as in  claim 20  further comprising means for automatic control of the operation of the system. 
     
     
       22. A system as in claim  21   18 wherein the means for automatic control is programmed to displace gel originally in the capillary by filling gel from the receptacle into the capillary. 
     
     
       23. An apparatus as in  claim 22  further comprising means for analyzing electrophoretically separated components of the sample. 
     
     
       24. An apparatus as in  claim 23  further comprising a cartridge for supporting the capillary. 
     
     
       25. A system as in claim  16   17 wherein the capillary has inner walls coated with a material that prevents bonding of the gel to the walls. 
     
     
       26. A system as in  claim 25  wherein the gel has a composition comprising up to 6% acrylamide and 0-5% crosslinker . 
     
     
       27. The capillary column as in  claim 12  wherein the material comprises phenyl groups, methyl groups or cyanopropyl groups. 
     
     
       28. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  filing means for filling the capillary with the polymerized gel filling;    
       
         means for introducing the sample into the filled capillary;  
       
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species; and    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel;    
       
         wherein the filing means and removing means flush spent gel and refill the capillary with fresh polymerized gel in a single step. 
       
     
     
       29. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  filling means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary; and    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel; and    
         g )  means for automatic control of operation of the apparatus;    
       
         wherein the filling means and removing means flush spent gel and refill the capillary with fresh polymerized gel in a single step. 
       
     
     
       30. A capillary electrophoresis system comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  a supply of sample;    
         d )  filling means for filling the capillary with the polymerized gel;    
         e )  means for introducing a sample into the capillary; and    
         f )  means for performing electrophoresis;    
       
         wherein the filling means also flushes spent gel and refills the capillary with fresh polymerized gel in a single step. 
       
     
     
       31. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species; and    
         f )  removing means for removing spent gel from to capillary so that it can be refilled with polymerized gel, the removing means comprising a rinsing device having a solution for expelling spent gel from the capillary.   
     
     
       32. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel, the removing means comprising a rinsing device having a solution for expelling spent gel from the capillary gel; and    
         g )  means for automatic control of operation of the apparatus.   
     
     
       33. A capillary electrophoresis system comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  a supply of sample;    
         d )  means for filling the capillary with the polymerized gel;    
         e )  means for introducing a sample into the capillary;    
         f )  means for performing electrophoresis; and    
         g )  a removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel, the removing means comprising a rinsing device having a solution for expelling gel from the capillary.   
     
     
       34. A system as in  claim 33 , further comprising means for automatic control of the operation of the capillary electrophoresis system. 
     
     
       35. A system as in  claim 34 , wherein the means for automatic control is programmed to flush the spent gel and refill the capillary with fresh polymerized gel in a single step. 
     
     
       36. A system as in  claim 34  wherein the means for automatic control is programmed to run the rinsing device following electrophoresis so as to displace the spent gel. 
     
     
       37. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel; and    
         g )  means for automatic control of operation of the apparatus;    
       
         wherein the polymerized gel and sample are contained in receptacles, and  
       
       
         wherein the apparatus further comprises a carrier for supporting the receptacles and means for positioning the carrier to place a respective receptacle in flow communication with the capillary. 
       
     
     
       38. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel; and    
         g )  means for automatic control of operation of the apparatus,    
       
         wherein the polymerized gel and sample are contained in receptacles and the apparatus further comprises a carrier for supporting the receptacles and means for positioning the carrier to place a respective receptacle in flow communication with the capillary, and  
       
       
         wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing. 
       
     
     
       39. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with polymerized gel; and    
         g )  means for automatic control of operation of the apparatus,    
       
         wherein the polymerized gel and sample are contained in receptacles and the apparatus further comprises a carrier for supporting the receptacles and means for positioning the carrier to place a respective receptacle in flow communication with the capillary, and  
       
       
         wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and  
       
       
         wherein the means for automatic control of the apparatus is programmed to displace the spent gel originally in the capillary by filling with polymerized gel from the receptacle into the capillary. 
       
     
     
       40. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
       
         removing means for removing spent gel from the capillary so that it can be refilled with a polymerized gel;  
       
         g )  means for automatic control of an operating system, and    
         h )  means for analyzing electrophoretically separated components of the sample,    
       
         wherein the polymerized gel and sample are contained in receptacles, and the apparatus further comprises a carrier for supporting the receptacles and means for positioning the carrier to place a respective receptacle in flow communication with the capillary, and  
       
       
         wherein the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and  
       
       
         wherein the means for automatic control is programmed to displace the spent gel originally in the capillary by filling with a polymerized gel from the receptacle into the capillary, and  
       
       
         further comprising means for analyzing electrophoretically separated components of the sample. 
       
     
     
       41. An apparatus for separating a sample into its molecular species comprising: 
         a )  a capillary and a cartridge for supporting the capillary;    
         b )  a supply of polymerized gel disposed in communication with one end of the capillary;    
         c )  means for filling the capillary with the polymerized gel;    
         d )  means for introducing the sample into the filled capillary;    
         e )  means for performing electrophoresis on the sample to separate the sample into its molecular species;    
         f )  removing means for removing spent gel from the capillary so that it can be refilled with a polymerized gel;    
         g )  means for automatic control of an operating system, and    
         h )  means for analyzing electrophoretically separated components of the sample,    
       
         wherein the polymerized gel and sample are contained in receptacles, and  
       
       
         wherein the system further comprises a carrier for supporting the receptacles and means for positioning the carrier to place a receptacle in flow communication with the capillary, and  
       
       
         the means for filling the capillary and introducing the sample comprises means for pressurizing the receptacles in order to effect gel filling and sample introducing, and  
       
       
         wherein the means for automatic control is programmed to displace the spent gel originally in the capillary by filling with polymerized gel from the receptacle into the capillary, and  
       
       
         wherein the apparatus further comprises a cartridge for supporting the capillary. 
       
     
     
       42. A gel-containing capillary column useful for electrophoresis comprising: 
       
         a capillary having a bore coated with a material having a property which prevents bonding of gel to the material; and  
       
       
         a crosslinked polymeric gel filling said bore, wherein the gel has a composition that allows it to be forced to fill the bore substantially without damage to gel structure which might otherwise significantly affect the electrophoretic performance of the gel. 
       
     
     
       43. The apparatus of  claim 2 ,  3 ,  4 ,  5 ,  8 ,  9 ,  28 ,  29 ,  31 ,  32 ,  37 ,  38 ,  39 ,  40 , or  41  wherein the means for filling the capillary with the polymerized gel refills the capillary with polymerized gel after the removing means removes spent gel from the capillary. 
     
     
       44. The system of  claim 17 ,  30  or  33  wherein the means for filling the capillary with the polymerized gel refills capillary with polymerized gel after the removing means removes spent gel from the capillary. 
     
     
       45. The apparatus of  claim 10  wherein the gel has a composition further comprising crosslinker in an amount up to  5 %. 
     
     
       46. The system of  claim 26  wherein the gel has a composition further comprising crosslinker in an amount up to  5 %.

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