Protein chromatography system

• US 6,344,172 B1
• Filed: 11/03/1994
• Issued: 02/05/2002
• Est. Priority Date: 09/30/1991
• Status: Expired due to Fees

• Current Assignee: Perseptive Biosystems Incorporated (Ecovyst, Inc.)
• Original Assignee: Perseptive Biosystems Incorporated (Ecovyst, Inc.)
• Inventors: Gordon, Neal F., Afeyan, Noubar
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Counts, Gary W.
• Application Number:
  US08/333,880
• Time in Patent Office: 2,651 Days
• Field of Search: 422/70, 422/81, 422/103, 210/656, 210/198.2, 210/96.1
• US Class Current: 422/70
• CPC Class Codes: B01D 15/1871 placed in series
• CPC Class Codes: B01D 15/325 Reversed phase
• CPC Class Codes: B01D 15/345 Perfusive chromatography
• CPC Class Codes: B01D 15/361 Ion-exchange
• CPC Class Codes: B01J 20/287 Non-polar phases; Reversed phases
• CPC Class Codes: B01J 41/05 in the strongly basic form
• CPC Class Codes: B01J 41/20 Anion exchangers for chromatographic processes
• CPC Class Codes: B03C 3/155 Filtration
• CPC Class Codes: G01N 2030/385 by switching valves
• CPC Class Codes: G01N 2030/524 structural properties
• CPC Class Codes: G01N 2030/625 by measuring reference material, e.g. carrier without sample
• CPC Class Codes: G01N 2030/8804 automated systems
• CPC Class Codes: G01N 2030/8813 biological materials
• CPC Class Codes: G01N 2030/8827 involving nucleic acids
• CPC Class Codes: G01N 2030/8831 involving peptides or proteins
• CPC Class Codes: G01N 2030/8872 impurities
• CPC Class Codes: G01N 30/08 using an enricher
• CPC Class Codes: G01N 30/24 Automatic injection systems
• CPC Class Codes: G01N 30/26 Conditioning of the fluid carrier; Flow patterns
• CPC Class Codes: G01N 30/40 using back flushing
• CPC Class Codes: G01N 30/461 : with serial coupling of separation columns
• CPC Class Codes: G01N 30/463 : for multidimensional chromatography
• CPC Class Codes: G01N 30/468 : involving switching between different column configurations
• CPC Class Codes: G01N 30/62 : Detectors specially adapted therefor
• CPC Class Codes: G01N 30/74 : Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J
• CPC Class Codes: G01N 30/88 : Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86

Abstract

The invention features an apparatus for the separation and analysis of proteins, which includes a sample input, a first liquid chromatography column, a multiport injection valve connecting the sample input to the column, a pump for providing variable pressure delivery of a solution to the column via the multiport valve, and a program for specifying a sequence of system control programs.

Claims

1. An apparatus for the separation of proteins in a sample, said apparatus comprising

sample solution input means,

a first liquid chromatography column,

at least one multiport injection valve connecting said sample solution input means to said column,

a second liquid chromatography column in communication with said multiport injection valve, said second column being operative successively with or alternatively to said first column,

at least one of said first and second columns being packed with a particulate chromatography material comprising perfusive particles which permit convective fluid transport both within and between the particles, or non-porous particles, thereby to confer on said column a chromatographically effective mobile phase transit time therethrough less than five minutes,

pump means for providing variable pressure delivery of solutions to said columns via said multiport valve,

plural solution reservoirs,

a mixing valve, connecting said solution reservoirs to said columns, operative to mix solutions from said reservoirs, and

program means for specifying a sequence of system control programs, the mixing of solution by said mixing valve, and the delivery of said mixed solution to said columns via said multiport injection valve.

2. The apparatus of claim 1, further comprising control means in communication with said pump means for controlling the pressure of delivery of said solution.

3. The apparatus of claim 1, further comprising

detector means for detecting and recording column output.

4. The apparatus of claim 3, further comprising template matching means for identifying a pattern of detected output data, said template matching means being operatively keyed to means for developing a control program for chromatographic separation.

5. An apparatus for the separation of proteins in a sample, comprising

a multiport mixing valve for mixing sample with one or more buffers to produce a sample mix,

plural liquid chromatography columns packed with a particulate chromatography material comprising perfusive particles which permit convective fluid transport both within and between the particles, or non-porous particles, thereby to confer on said columns a chromatographically effective liquid phase transit time therethrough less than five minutes, each said column comprising a first and a second end,

a multiport injection valve in communication with said sample mixing valve and said first end of each of said liquid chromatography columns,

an output system comprising an output signal recording system and an output sample collection system, wherein said output system is in communication with said second end of each of said columns, and

control means for operating said multiport injection valve to successively and alternately apply a mixed solution from the mixing valve to one of said respective columns in coordination with operation of said output system to run a sequence of separations for the preparation or analysis of a protein.

6. The apparatus of claim 5, further comprising

a sample input system comprising plural solution reservoirs and a sample reservoir.

7. The apparatus of claim 5 wherein said control means further comprises

switching means for alternatively utilizing one of said plural liquid chromatography columns while cleaning another, thus providing a substantially continuous operating sequence of outputs from successively utilized columns.

8. The apparatus of claim 5 further comprising

program means for specifying a sequence of separation process control programs to be successively run during operation.

9. The apparatus of claim 8 wherein said program means specifies a separation program in which first and second columns are utilized successively for separating proteins in said sample.

10. The apparatus of claim 9 wherein one said column comprises an ion exchange chromatography matrix.

11. The apparatus of claim 9 wherein one said column comprises a reverse phase chromatography matrix.

12. The apparatus of claim 8 wherein said program specifies substantially continuous preparation of a separated sample in said first column and intermittent analysis of said first column output via said second column.

13. The apparatus of claim 5 wherein at least one of said plural columns is removable and replaceable by a different column.

14. An apparatus for the quantitative detection of proteins in a sample, comprising

first and second multiport valves, each said valve comprising a sample loop which holds a defined volume of sample and connects two ports of said valve;

a liquid chromatography column in communication with said valve, said column being packed with a particulate chromatography material comprising perfusive particles which permit convective fluid transport both within and between the particles, or nonporous particles, thereby to confer on said column a chromatographically effective mobile phase transit time therethrough less than five minutes;

a sample feed line in communication with each said valve;

detector means in communication with said column for detecting output; and

control means for operating said multiport valves to switch between

a collection line comprising said sample feed line, wherein plural sample volumes are introduced into said sample loops and

a detection line comprising said chromatography column, wherein one sample volume is passed directly through said detector means and another is passed through said column and said detector means.

15. The apparatus of claim 14, wherein said collection line comprises in successive order

(a) a first sample loop connecting within said first valve a first port and a second port,

(b) said sample feed line, said feed line connecting an exit port in said first valve with an entry port of said second valve, and

(c) a second sample loop connecting within said second valve an entry port and an exit port.

16. The apparatus of claim 14 wherein said detection line comprises in successive order

(a) a first sample loop connecting within said first valve a first port and a third port,

(b) said chromatography column, said column connecting said third port of said first valve with a third port of said second valve, and

(c) a second sample loop connecting within said second valve said third port to said detector means.

17. The apparatus of claim 14 wherein said apparatus further comprises a third multiport valve and said chromatography column is in communication with said first and second valves through said third valve.