Automated cell processing instruments comprising reagent cartridges

• US 10,406,525 B1
• Filed: 06/25/2019
• Issued: 09/10/2019
• Est. Priority Date: 04/13/2018
• Status: Active Grant

• Current Assignee: Inscripta, Inc.
• Original Assignee: Inscripta, Inc.
• Inventors: Masquelier, Don, Belgrader, Phillip, Van Hatten, Brian, Bernate, Jorge, Chabansky, Bruce
• Primary Examiner(s): Hobbs, Michael L
• Application Number:
  US16/451,602
• Time in Patent Office: 77 Days
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/04 Closures and closing means
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 3/527 for a plurality of reagents
• CPC Class Codes: B01L 7/00 Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• CPC Class Codes: B01L 7/02 Water baths; Sand baths; Air baths
• CPC Class Codes: C12M 23/06 Tubular C12M23/08, C12M23/16 take precedence
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/18 Heat exchange systems, e.g. heat jackets or outer envelopes
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves

Abstract

The present disclosure provides a reagent cartridge configured for use in an automated multi-module cell processing environment.

Claims

1. A kit comprising a reagent cartridge for an automated multi-module cell processing instrument, wherein the reagent cartridge comprises:

a thermal block with block reservoirs defined therein;

a cover disposed upon the thermal block with cover reservoirs defined therein, wherein the cover reservoirs and block reservoirs are coincident and together define reservoirs in the reagent cartridge;

an insert comprising a flow-through electroporation device;

a script readable by a processor;

at least two additional inserts configured to reside within the reservoirs in the reagent cartridge; and

reagents dispensed in one or more of the reagent reservoirs.

2. The kit of claim 1, wherein the one or more reagent reservoirs with reagents dispensed therein is sealed.

3. The kit of claim 1, further comprising a top cover configured to seal the cover reservoirs.

4. The kit of claim 1, further comprising a rotating growth vial with media and cells dispensed therein.

5. The kit of claim 4, wherein the cells are bacterial cells.

6. The kit of claim 5, wherein the cells are E. coli cells.

7. The kit of claim 4, wherein the cells are yeast cells.

8. The kit of claim 7, wherein the cells are S. cerevisiae cells.

9. The kit of claim 4, wherein the cells are plant cells.

10. The kit of claim 4, wherein the cells are animal cells.

11. The kit of claim 10, wherein the cells are mammalian cells.

12. The kit of claim 11, wherein the cells are human cells.

13. The kit of claim 1, wherein one of the reagents dispensed in the one or more reagent reservoirs comprises an antibiotic.

14. The kit of claim 1, wherein one of the reagents dispensed in the one or more reagent reservoirs comprises a nucleic acid-guided nuclease.

15. The kit of claim 14, wherein the nucleic acid-guided nuclease is selected from Cas 9, Cas 12/CpfI, MAD2, or MAD7.

16. The kit of claim 15, wherein the nucleic acid-guided nuclease is MAD7.

17. The kit of claim 1, wherein the flow-through electroporation device comprises:

an inlet and inlet channel for introduction of a cell sample to the flow-through electroporation device;

an outlet and outlet channel for exit of the cell sample from the flow-through electroporation device;

a flow channel intersecting and positioned between the inlet channel and outlet channel with a constriction in the flow channel positioned between the inlet channel and outlet channel; and

two or more electrodes, wherein at least one of the electrodes is positioned in the flow channel between the intersection of the flow channel with the first inlet channel and the constriction in the flow channel and at least one of the electrodes is positioned between the constriction in the flow channel and the intersection of the flow channel with the outlet channel, and wherein the two or more electrodes are in fluid communication with the cell sample in the flow channel but are not in the flow path of the cell sample in the flow channel, and wherein the two or more electrodes are configured to apply an electric pulse or electric pulses to a cell sample.

20. The kit of claim 1, further comprising a rotating growth vial with media and cells dispensed therein.

18. A kit comprising a reagent cartridge for an automated multi-module cell processing instrument, wherein the reagent cartridge comprises:

a thermal block with block reservoirs defined therein;

a cover disposed upon the thermal block with cover reservoirs defined therein, wherein the cover reservoirs and block reservoirs are coincident and together define reservoirs in the reagent cartridge;

an insert comprising a flow-through electroporation device;

a script readable by a processor;

at least two additional inserts configured to reside within the reservoirs in the reagent cartridge,

wherein a nuclease is dispensed in one of the at least two additional inserts, and an antibiotic is dispensed in another of the at least two additional inserts.

19. The kit of claim 18, wherein the flow-through electroporation device comprises:

an inlet and inlet channel for introduction of a cell sample to the flow-through electroporation device;

an outlet and outlet channel for exit of the cell sample from the flow-through electroporation device;

a flow channel intersecting and positioned between the inlet channel and outlet channel with a constriction in the flow channel positioned between the inlet channel and outlet channel; and

two or more electrodes, wherein at least one of the electrodes is positioned in the flow channel between the intersection of the flow channel with the first inlet channel and the constriction in the flow channel and at least one of the electrodes is positioned between the constriction in the flow channel and the intersection of the flow channel with the outlet channel, and wherein the two or more electrodes are in fluid communication with the cell sample in the flow channel but are not in the flow path of the cell sample in the flow channel, and wherein the two or more electrodes are configured to apply an electric pulse or electric pulses to a cell sample.