Chemical sample treatment system and cassette, and methods for effecting multistep treatment process

• US 6,039,924 A
• Filed: 09/03/1998
• Issued: 03/21/2000
• Est. Priority Date: 07/09/1996
• Status: Expired due to Fees

• Current Assignee: Amersham Pharmacia Biotech AB
• Original Assignee: Amersham Pharmacia Biotech AB
• Inventors: Horn, Marcus J.
• Primary Examiner(s): NOLAND, THOMAS
• Application Number:
  US09/146,160
• Time in Patent Office: 565 Days
• Field of Search: 422/67, 422/63, 422/70, 422/101, 422/102, 436/54, 141/130, 73/61.55
• US Class Current: 422/67
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: B01L 3/50855 using modular assemblies of strips or of individual wells
• CPC Class Codes: B01L 9/06 Test-tube stands; Test-tube holders
• CPC Class Codes: G01N 2030/027 Liquid chromatography
• CPC Class Codes: G01N 2030/8804 automated systems
• CPC Class Codes: G01N 2030/8831 involving peptides or proteins
• CPC Class Codes: G01N 2030/8881 Modular construction, specially adapted therefor
• CPC Class Codes: G01N 2035/00752 bar codes
• CPC Class Codes: G01N 30/24 Automatic injection systems
• CPC Class Codes: G01N 30/36 in high pressure liquid systems
• CPC Class Codes: G01N 30/466 with separation columns in parallel
• CPC Class Codes: G01N 30/6043 in parallel
• 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
• CPC Class Codes: G01N 35/00732 Identification of carriers, materials or components in automatic analysers
• CPC Class Codes: Y10T 436/119163 with aspirator of claimed structure

Abstract

This invention is directed to a cassette chemical immobilization and treatment system and method for enabling the performance of various complex chemistries with minimal human intervention, near-zero dead volume, and flow-through protocols pursuant to a predetermined instruction set encoded on a multiple-address chemical treatment cassette assembly. The cassette assembly having a high-pressure interface capability to permit direct insertion of the mini-column into a high-pressure solvent line for use as a support column for HPLC analysis. A sample loader loads analyte samples either singly or simultaneously into the multiple, addressable mini-columns without the need to load the sample funnel/column assembly into a separate reaction chamber. The invention system reads the machine readable instruction code and performs chemistries on the analyte, including mixing solvents, reactants and buffers; detaching and shuttling the analyte back and forth performing complex chemistries either on or off-column; solvation of powdered or lyophillized reagents; and performing in-line, on-board analysis. These protocols are performed automatically pursuant to instructions contained on a bar code, without having to remove the cassette or interface from the CTS or otherwise require human intervention. The same chemistries may be performed on all sample mini-column addresses, or a separate completely independent set of protocols may be defined for each address, or for each block of addresses pursuant to the instructions contained in the machine readable code on the cassette.

Claims

1. A chemical treatment system for sequential, uninterrupted treatment of less than milligram quantities of a plurality of chemical samples, comprising in operative combination:

a) a chemical treatment cassette for retaining a plurality of chemical samples for chemical treatment, said chemical treatment cassette further comprising;

i) a funnel assembly comprising a plurality of integrated, individually addressable chemical solution sampling funnels for receiving chemical sample solutions, each of said sampling funnels having a first, open mouth end for introduction of a chemical sample solution, and a tapered, second conical end having a through-hole at the apex of said conical end, said sampling funnels being arranged in an array to permit individual addressing for chemical treatment;

ii) a plurality of sample mini-columns for retaining preselected chemical samples, each of said mini-columns having a centrally disposed chamber extending longitudinally through said mini-column, at least one of said mini-columns having a material selected from at least one of a solid support, dry reagent, dry sample, liquid reagent, liquid sample, and porous frits disposed in said central chamber, said chamber terminating into a first open end, said first open end having a flared, tapered inlet port, said inlet port being in flow-through, physical connection with said through-hole in said conical second end of said funnel, said connection resulting in smooth, continuous transition of the taper from said conical second end of said funnel to said taper of said inlet port, said chamber terminating into a second open end, said second open end having a flared, tapered outlet port, said outlet port being in flow-through communication with said inlet port, said first open end of each of said funnels and said tapered conical second end of each of said funnels and permitting localized access by a compressive fit interface to said mini-column inlet port for near-zero dead volume flow-through communication between said interface and said mini-columns;

iii) at least one reagent well for retaining preselected reagents for reaction with said preselected sample retained in said min-columns, said reagent wells comprising a through-bore parallel to said longitudinal axis of said mini-column, said through-bore terminating in a first end having a flared, tapered inlet port, and terminating in a second end having a flared, outlet port said reagent well inlet port being in flow-through communication with said reagent well outlet port; and

iv) a machine readable code disposed on said chemical treatment cassette for identifying the chemistry protocols to be performed on the retained samples in said mini-columns;

b) a sample loading station for loading sample solutions in said sample funnels, said sample loading station further comprising;

i) a cassette alignment mechanism for aligning said chemical treatment cassette to a predetermined X-Y position;

ii) a sealing gasket sealing said first end of said sampling funnel when said sample funnel is located at said predetermined X-Y position;

iii) a pressure cap having an annular shoulder and center plunger section for press-fit insertion into the mouth of said sample funnel to define a funnel headspace, said shoulder engaging said annular gasket to limit the extent of insertion of said center plunger, said pressure cap having a through-bore gas inlet for introduction of pressurized gas into said funnel headspace to force said sample solution into said inlet port of said mini-column;

c) a chemical treatment station for performing predetermined chemistries on a plurality of preselected retained chemical samples located in said chemical treatment cassette, said chemical treatment station further comprising;

i) a code reader for reading said machine readable code disposed on said chemical treatment cassette to identify the specific chemistry protocols to be performed on said retained chemical samples;

ii) a scanner to identify mini-column addresses wherein said central chamber of said mini-column is populated;

iii) a micro-controller interface for communicating from said code reader to a micro-controller said identified specific chemistry protocols, said micro-controller accessing specific process algorithms pursuant to the specific identified chemistry protocols, said process algorithms defining and communicating to said micro-controller interface stepwise micro-controller control instructions to enable execution of said identified specific chemistries on said pre-selected chemical samples at said populated mini-column addresses;

iv) a cassette/treatment station interface for enabling near-zero dead volume flow-through communication of said chemical treatment cassette with said chemical treatment station, said cassette/treatment station interface comprising;

a cassette interface dock for precision alignment of said chemical treatment cassette,

a first nozzle array comprising a plurality of nozzles spatially arrayed to be congruous with said mini-column inlet ports and reagent well inlet ports for flow-through localized interfacing with said mini-column inlet ports and said reagent well inlet ports,

a second nozzle array having a plurality of nozzles spatially arrayed to be congruous with said mini-column outlet ports and reagent well outlet ports for flow through communication with said mini-column outlet ports and said reagent well outlet ports,

said first nozzle array positioned above said cassette interface dock, said nozzles in said first array having nozzles tips extending downward towards said cassette interface dock,

said second nozzle array position below said cassette interface dock, said nozzles in said second nozzle array having nozzle tips extending upwards to said cassette interface dock, and

said nozzles in said first array and said nozzles in said second array establishing said flow-through localized interface by first aligning said chemical treatment cassette in said cassette interface dock and by compression fitting said nozzle tips in said first nozzle array against said tapered opening of said mini-column inlet ports and said reagent well inlet ports, and by compression fitting said nozzle tips in said second nozzle array against said tapered opening of said mini-column outlet ports and said reagent well outlet ports;

v) a chemical delivery subsystem for providing automatic flow through chemistries on said preselected chemical samples further comprising;

a solvent bank providing a plurality of reagents and solvents to said reagent wells for solubilizing said reagents retained therein and for reaction with said chemical samples pursuant to said identified specific chemistry protocols,

a rotary valve array further comprising a mini-column inlet port rotary valve array for individual addressing of mini-column inlet ports, a mini-column outlet port rotary valve array for individual addressing of mini-column outlet ports, said mini-column inlet port rotary valve array and said mini-column outlet port rotary valve array being synchronized to a single mini-column address to permit active interface with only one mini-column inlet and outlet port at a time, and a reagent well inlet port rotary valve array for individual addressing of reagent well inlet ports, a reagent well outlet port rotary valve array for individual addressing of reagent well outlet ports, said reagent well inlet port rotary valve array and said reagent well outlet port rotary valve array being synchronized to a single reagent well address to permit active interface with only one reagent well inlet and outlet port at a time,

a plurality of accumulators for temporary storage of solvents and reagents, and temporary storage of detached chemical sample to enable chemistries to be performed on said detached samples, and to enable chemistries not including said detached samples to be performed in said mini-column,

a mixer/diluter reservoir in communication with said solvent bank, reagent wells, and mini-columns for temporary retention of at least one of a solvent, reagent, sample, and mixtures thereof,

a pneumatic pressurizing system for bi-directional transfer of solvents, reagents, and chemical samples to and from said reagent wells, said mini-columns, said accumulators, and said mixer/diluter, and for supplying inert gas to said chemical treatment delivery subsystem, and

a valve for integration of an analyzer subsystem into said chemical delivery subsystem to permit automatic analysis of the sample retained in said mini-columns without removal of said mini-columns from said chemical treatment cassette and without disengaging said cassette/treatment station interface.

2. A chemical treatment system as in claim 1 wherein said chemical treatment cassette further comprises an alignment mechanism for aligning said mini-columns such that the longitudinal axis of said mini columns is coincident with the longitudinal axis of said sample funnels.

3. A chemical treatment system as in claim 1 wherein a solid support material is disposed in said central chamber of said mini-column, said solid support material being treated to immobilize said chemical sample, said solid support being retained in said mini-columns by porous frits disposed in said central chamber at said inlet port end and at said outlet port end.

4. A chemical treatment system as in claim 1 wherein said mini-columns are integral to said funnel assembly to provide a unitary, one-piece funnel/mini-column assembly.

5. A chemical treatment system for treatment of less than milligram quantities of a chemical sample, comprising in operative combination:

a) a chemical sample retaining means for holding a plurality of chemical samples for chemical treatment, said chemical sample retaining means further comprising;

i) a sample funneling means for receiving a plurality of chemical sample solutions, said sample funneling means permitting individual addressing of each of said plurality of chemical sampling solutions for chemical treatment;

ii) means for retaining an individual preselected chemical sample, said individual sample retaining means being in flow-through, physical connection with said funneling means, said means permitting near-zero dead volume compression fit with an interface means;

iii) at least one reagent retaining means for retaining preselected reagents, said reagent retaining means permitting flow-through communication there through;

iv) a machine readable code disposed on said chemical sample retaining means for identifying the chemistry protocols to be performed on the retained samples in said individual sample retaining means;

b) a sample loading means for loading sample solutions in said sample funneling means, said sample loading means further comprising;

i) an alignment means for aligning said chemical sample retaining means to a predetermined X-Y position;

ii) a sealing gasket on said sample funneling means on said funneling means;

iii) a pressure cap means for introduction of pressurized gas into said sample funneling means and for forcing a sample solution through said individual sample retaining means;

c) a chemical treatment station for performing predetermined chemistries on a plurality of preselected retained chemical samples located in said chemical sample retaining means, said chemical treatment station further comprising;

i) a code reader for reading said machine readable code disposed on said chemical treatment cassette to identify the specific chemistry protocols to be performed on said retained chemical samples;

ii) a scanner to identify populated mini-column addresses;

iii) a micro-controller interface means for communicating from said code reader to a micro-controller said identified specific chemistry protocols, said micro-controller accessing specific process algorithms pursuant to the specific identified chemistry protocols, said process algorithms defining and communicating to said micro-controller interface step-wise micro-controller control instructions to enable execution of said identified specific chemistries on said pre-selected chemical samples at said populated mini-column addresses;

iv) a chemical cassette/treatment interface means for enabling flow-through localized communication of said chemical sample retaining means with said chemical treatment station, said interface means comprising;

a docking means for precision alignment of said chemical sample retaining means,

an interface means for establishing flow-through localized interface between said chemical treatment station and said chemical sample retaining means;

v) a chemical delivery subsystem for providing automatic flow through chemistries on said preselected chemical samples further comprising;

a solvent storage means for providing a plurality of reagents and solvents to said reagent retention means for solubilizing said reagents retained therein and to said chemical sample retaining means for treatment of said chemical samples in said individual sample retaining means pursuant to said identified specific chemistry protocols,

a valve means for individual addressing of said individual sample retaining means and for individual addressing of said reagent retention means,

a plurality of accumulator means for temporary storage of solvents and reagents, and temporary storage of detached chemical sample to enable chemistries to be performed on said detached samples, and to enable chemistries not including said detached samples as a reactant to be performed in said individual sample retaining means,

a mixer/diluter reservoir in communication with said solvent bank, reagent wells, and mini-columns for temporary retention of at least one of a solvent, reagent, sample, and mixtures thereof,

a pneumatic pressurizing system for bi-directional transfer of solvents, reagents, and chemical samples to and from said reagent wells, said mini-columns, said accumulators, and said mixer/diluter, and for supplying inert gas to said chemical treatment delivery subsystem, and

valve means for integration of an analyzer subsystem into said chemical delivery subsystem to permit automatic analysis of the sample retained in said mini-columns without removal of said mini-columns from said chemical treatment cassette and without disengaging, said cassette/treatment station interface.

6. A chemical treatment system for sequential, uninterrupted treatment of less than milligram quantities of a plurality of chemical samples, comprising in operative combination:

a) a chemical treatment cassette for retaining a plurality of chemical samples for chemical treatment, said chemical treatment cassette further comprising;

i) a funnel assembly comprising a plurality of integrated, individually addressable chemical solution sampling funnels for receiving chemical sample solutions, each of said sampling funnels having a first, open mouth end for introduction of a chemical sample solution, and a tapered, second conical end having a through-hole at the apex of said conical end, said sampling funnels being arranged in an array to permit individual addressing for chemical treatment;

ii) a plurality of sample mini-columns for retaining preselected chemical samples, each of said mini-columns having a centrally disposed chamber extending longitudinally through said mini-column, at least one of said mini-columns having a material selected from at least one of a solid support, dry reagent, dry sample, liquid reagent, liquid sample, and porous frits disposed in said central chamber, said chamber terminating into a first open end, said first open end having a flared, tapered inlet port, said inlet port being in flow-through, physical connection with said through-hole in said conical second end of said funnel, said connection resulting in smooth, continuous transition of the taper from said conical second end of said funnel to said taper of said inlet port, said chamber terminating into a second open end, said second open end having a flared, tapered outlet port, said outlet port being in flow-through communication with said inlet port, said first open end of each of said funnels and said tapered conical second end of each of said funnels and permitting localized access by a compressive fit interface to said mini-column inlet port for near-zero dead volume flow-through communication between said interface and said mini-columns;

iii) at least one reagent well for retaining preselected reagents for reaction with said preselected sample retained in said min-columns, said reagent wells comprising a through-bore parallel to said longitudinal axis of said mini-column, said through-bore terminating in a first end having a flared, tapered inlet port, and terminating in a second end having a flared, outlet port said reagent well inlet port being in flow-through communication with said reagent well outlet port; and

iv) a machine readable code disposed on said chemical treatment cassette for identifying the chemistry protocols to be performed on the retained samples in said mini-columns;

b) a sample loading station for loading sample solutions in said sample funnels, said sample loading station including an X-Y positioner for aligning said chemical treatment cassette, and a pressure cap having an annular shoulder and center plunger section for press-fit insertion into the mouth of said sample funnel to define a funnel headspace, said shoulder limiting the extent of insertion of said center plunger, said pressure cap having a though-bore gas inlet for introduction of pressurized gas into said funnel headspace to force said sample solution into said inlet port of said mini-column;

c) a chemical treatment station for performing predetermined chemistries on a plurality of preselected retained chemical samples located in said chemical treatment cassette, said chemical treatment station further comprising;

i) a code reader for reading said machine readable code disposed on said chemical treatment cassette to identify the specific chemistry protocols to be performed on said retained chemical samples;

ii) a scanner to identify mini-column addresses wherein said central chamber of said mini-column is populated;

iii) a micro-controller interface for communicating from said code reader to a micro-controller said identified specific chemistry protocols, said micro-controller accessing specific process algorithms pursuant to the specific identified chemistry protocols, said process algorithms defining and communicating to said micro-controller interface stepwise micro-controller control instructions to enable execution of said identified specific chemistries on said pre-selected chemical samples at said populated mini-column addresses;

iv) a cassette/treatment station interface for enabling near-zero dead volume flow-through communication of said chemical treatment cassette with said chemical treatment station, said cassette/treatment station interface comprising;

a cassette interface dock for precision alignment of said chemical treatment cassette,

a first nozzle array comprising a plurality of nozzles spatially arrayed to be congruous with said mini-column inlet ports and reagent well inlet ports for flow-through localized interfacing with said mini-column inlet ports and said reagent well inlet ports,

a second nozzle array having a plurality of nozzles spatially arrayed to be congruous with said mini-column outlet ports and reagent well outlet ports for flow through communication with said mini-column outlet ports and said reagent well outlet ports,

said first nozzle array positioned above said cassette interface dock, said nozzles in said first array having nozzles tips extending downward towards said cassette interface dock,

said second nozzle array position below said cassette interface dock, said nozzles in said second nozzle array having nozzle tips extending upwards to said cassette interface dock, and

said nozzles in said first array and said nozzles in said second array establishing said flow-through localized interface by first aligning said chemical treatment cassette in said cassette interface dock and by compression fitting said nozzle tips in said first nozzle array against said tapered opening of said mini-column inlet ports and said reagent well inlet ports, and by compression fitting said nozzle tips in said second nozzle array against said tapered opening of said mini-column outlet ports and said reagent well outlet ports;

v) a chemical delivery subsystem for providing automatic flow through chemistries on said preselected chemical samples further comprising;

a solvent bank providing a plurality of reagents and solvents to said reagent wells for solubilizing said reagents retained therein and for reaction with said chemical samples pursuant to said identified specific chemistry protocols,

a rotary valve array further comprising a mini-column inlet port rotary valve array for individual addressing of mini-column inlet ports, a mini-column outlet port rotary valve array for individual addressing of mini-column outlet ports, said mini-column inlet port rotary valve array and said mini-column outlet port rotary valve array being synchronized to a single mini-column address to permit active interface with only one mini-column inlet and outlet port at a time, and a reagent well inlet port rotary valve array for individual addressing of reagent well inlet ports, a reagent well outlet port rotary valve array for individual addressing of reagent well outlet ports, said reagent well inlet port rotary valve array and said reagent well outlet port rotary valve array being synchronized to a single reagent well address to permit active interface with only one reagent well inlet and outlet port at a time,

a plurality of accumulators for temporary storage of solvents and reagents, and temporary storage of detached chemical sample to enable chemistries to be performed on said detached samples, and to enable chemistries not including said detached samples to be performed in said mini-column,

a mixer/diluter reservoir in communication with said solvent bank, reagent wells, and mini-columns for temporary retention of at least one of a solvent, reagent, sample, and mixtures thereof,

a pneumatic pressurizing system for bi-directional transfer of solvents, reagents, and chemical samples to and from said reagent wells, said mini-columns, said accumulators, and said mixer/diluter, and for supplying inert gas to said chemical treatment delivery subsystem, and

a valve for integration of an analyzer subsystem into said chemical delivery subsystem to permit automatic analysis of the sample retained in said mini-columns without removal of said mini-columns from said chemical treatment cassette and without disengaging said cassette/treatment station interface.

7. A chemical treatment system as in claim 6 wherein said chemical treatment cassette further comprises an alignment mechanism for aligning said mini-columns such that the longitudinal axis of said mini columns is coincident with the longitudinal axis of said sample funnels.

8. A chemical treatment system as in claim 6 wherein a solid support material is disposed in said central chamber of said mini-column, said solid support material being treated to immobilize said chemical sample, said solid support being retained in said mini-columns by porous frits disposed in said central chamber at said inlet port end and at said outlet port end.

9. A chemical treatment system as in claim 6 wherein said mini-columns are integral to said funnel assembly to provide a unitary, one-piece funnel/mini-column assembly.

10. A chemical treatment system for treatment of less than milligram quantities of a chemical sample, comprising in operative combination:

a) a chemical sample retainer for holding a plurality of chemical samples for chemical treatment, said chemical sample retainer further comprising;

i) a sample funnel for receiving a plurality of chemical sample solutions, said sample funnel permitting individual addressing of each of said plurality of chemical sampling solutions for chemical treatment;

ii) a retainer for an individual preselected chemical sample, said retainer being in flow-through, physical connection with said funnel, and providing near-zero dead volume compression fit;

iii) at least one reagent retainer for preselected reagents, said reagent retainer permitting flow-through communication therethrough;

iv) a machine readable code disposed on said chemical sample retainer for identifying the chemistry protocols to be performed on the retained samples in said individual sample retainer;

b) a sample loading mechanism for loading sample solutions in said sample funnel, and including an X-Y positioner for aligning said chemical sample retainer to a predetermined X-Y position, a sealing gasket, and a pressure cap for introduction of pressurized gas into said sample funnel and for forcing a sample solution through said individual sample retainer;

c) a chemical treatment station for performing predetermined chemistries on a plurality of preselected retained chemical samples located in said chemical sample retainer, said chemical treatment station further including;

i) a code reader for reading said machine readable code disposed on said chemical treatment cassette to identify the specific chemistry protocols to be performed on said retained chemical samples;

ii) a scanner to identify populated mini-column addresses;

iii) a micro-controller interface for communicating from said code reader to a micro-controller said identified specific chemistry protocols, said micro-controller accessing specific process algorithms pursuant to the specific identified chemistry protocols, said process algorithms defining and communicating to said micro-controller interface step-wise micro-controller control instructions to enable execution of said identified specific chemistries on said pre-selected chemical samples at said populated mini-column addresses;

iv) a chemical cassette/treatment interface for enabling flow-through localized communication of said chemical sample retainer with said chemical treatment station, said interface including a docking mechanism for precision alignment of said chemical sample retainer, and a flow-through localized interface between said chemical treatment station and slid chemical sample retainer;

v) a chemical delivery subsystem for providing automatic flow through chemistries on said preselected chemical samples, the chemical delivery subsystem including solvent storage for a plurality of reagents and solvents to said reagent retainer for solubilizing said reagents retained therein and to said chemical sample retainer for treatment of said chemical samples in said individual sample retainer pursuant to said identified specific chemistry protocols, a valve for individual addressing of said individual sample retainer and for individual addressing of said reagent retainer, a plurality of accumulators for temporary storage of solvents and reagents, and temporary storage of detached chemical sample to enable chemistries to be performed on said detached samples, and to enable chemistries not including said detached samples as a reactant to be performed in said individual sample retainer, a mixer/diluter reservoir in communication with said solvent bank, reagent wells, and mini-columns for temporary retention of at least one of a solvent, reagent, sample, and mixtures thereof, a pneumatic pressurizing system for bi-directional transfer of solvents, reagents, and chemical samples to and from said reagent wells, said mini-columns, said accumulators, and said mixer/diluter, and for supplying inert gas to said chemical treatment delivery subsystem, and a valve integrating of an analyzer subsystem into said chemical delivery subsystem to permit automatic analysis of the sample retained in said mini-columns without removal of said mini-columns from said chemical treatment cassette and without disengaging said cassette/treatment station interface.