Continuous automated perfusion culture analysis system (CAPCAS) and applications of same

Patent metadata

Field	Value
Publication number	US-11447734-B2
Application number	US-202217578966-A
Country	US
Kind code	B2
Filing date	Jan 19, 2022
Priority date	Jun 28, 2019
Publication date	Sep 20, 2022
Grant date	Sep 20, 2022

Abstract

A continuous automated perfusion culture analysis system (CAPCAS) comprises one or more fluidic systems configured to operate large numbers of biodevices in parallel. Each fluidic system comprises an input reservoir plate for receiving media; a biodevice plate comprising an array of biodevices fluidically coupled to the input reservoir plate, configured such that each biodevice has independent media delivery, fluid removal, stirring, and gas control, and each biodevice is capable of continuously receiving the media from the input reservoir plate; and an output plate fluidically coupled to the biodevice plate for real-time analysis and sampling. The operations of the CAPCAS are automated and computer-controlled wirelessly. The CAPCAS can also be used for abiotic and biotic chemical synthesis processes.

First claim

What is claimed is:

1. A continuous automated perfusion culture analysis system (CAPCAS), comprising: one or more fluidic systems configured to operate large numbers of biodevices in parallel, wherein each fluidic system comprises:
an input reservoir plate for receiving media;
a biodevice plate comprising an array of biodevices fluidically coupled to the input reservoir plate, configured such that each biodevice has independent media delivery, fluid removal, and gas control, and each biodevice is capable of continuously receiving the media from the input reservoir plate;
an output plate fluidically coupled to the biodevice plate for real-time analysis and sampling; and
at least one microformulator fluidically coupled to the input reservoir plate for providing the media to the input reservoir plate, wherein each microformulator comprises:
a plurality of feedstock solution reservoirs;
at least one input selector valve (V1) fluidically coupled to the plurality of feedstock solution reservoirs to select at least one feedstock reservoir;
at least one output director valve (V2) fluidically coupled to the input reservoir plate; and
at least one pump (P1) fluidically coupled between the at least one input selector valve and the at least one output director valve for withdrawing fluid from the selected feedstock solution reservoir through the at least one input selector valve and delivering it to the input reservoir plate through the at least one output director valve.

2. The CAPCAS of claim 1 , wherein the at least one input selector valve is a multichannel input selector valve, the at least one pump is a single-channel pump, and the at least one output director valve is a multichannel output director valve.

3. The CAPCAS of claim 1 , wherein the at least one input selector valve is configured to select different feedstock solution reservoirs at different periods of time.

4. The CAPCAS of claim 1 , wherein the at least one pump is driven such that the fluid of the selected feedstock solution reservoir outputs from the at least one output director valve at a predetermined flow rate.

5. The CAPCAS of claim 4 , wherein the predetermined flow rate varies with time.

6. The CAPCAS of claim 1 , wherein through a sequence of selecting the plurality of reservoirs by the at least one input selector valve and pump speed and duration actuations of the at least one pump, the media is provided to have a different time-varying perfusion mixture for each biodevice.

7. The CAPCAS of claim 1 , wherein each microformulator further includes a single-channel optical sensing module coupled between the at least one pump and the at least one output director valve for tracking an intentionally injected bubble for measurement of flow rate, or identifying when a reservoir is emptied.

8. A continuous automated perfusion culture analysis system (CAPCAS), comprising: one or more fluidic systems configured to operate large numbers of biodevices in parallel, wherein each fluidic system comprises:
an input reservoir plate for receiving media;
a biodevice plate comprising an array of biodevices fluidically coupled to the input reservoir plate, configured such that each biodevice has independent media delivery, fluid removal, and gas control, and each biodevice is capable of continuously receiving the media from the input reservoir plate;
an output plate fluidically coupled to the biodevice plate for real-time analysis and sampling; and
a biodevice media delivering means fluidically coupled between the input reservoir plate and the biodevice plate for continuous delivery of the media from the input reservoir plate to each biodevice.

9. The CAPCAS of claim 8 , wherein the input reservoir plate has two sets of media ports, and wherein the biodevice media delivering means comprises two multichannel pumps (P2, P3), each multichannel pump is fluidically coupled between a respective set of the media ports and the biodevice plate, such that one set is refillable while the other set is being delivered by a corresponding pump to each biodevice in the biodevice plate, thereby providing uninterrupted perfusion.

10. The CAPCAS of claim 9 , further comprising a biodevice media collecting means fluidically coupled between the biodevice plate, and the output plate and an analyzer for real-time analysis and sampling.

11. The CAPCAS of claim 10 , wherein the biodevice media collecting means comprises first and second multichannel pumps (P4, P5), and at least one output valve (V3), wherein the first multichannel pump (P4) is fluidically coupled between the biodevice plate and the at least one multimode output valve (V3), the second multichannel pump (P5) is fluidically coupled between the biodevice plate and the output plate, and the least one multimode output valve (V3) is fluidically coupled between the first multichannel pump (P4) and the analyzer.

12. The CAPCAS of claim 11 , wherein the second multichannel pump (P5) operates independent of the first multichannel pump (P4) to deliver the effluent from each biodevice to a separate well in the output plate for off-line transcriptomic or other off-line analysis.

13. The CAPCAS of claim 11 , wherein the at least one multimode output valve is configured to either divert effluent from each biodevice to the analyzer.

14. The CAPCAS of claim 11 , wherein the at least one multimode output valve is configured to divert, when one biodevice is being sampled, the media being pumped from the other biodevices to waste, with each biodevice being sampled serially.

15. The CAPCAS of claim 11 , wherein the at least one multimode output valve is configured to divert the effluent from all the biodevices to waste to ensure continuous perfusion when no sample is needed or the output plate is removed after bulk sample collection.

16. The CAPCAS of claim 11 , wherein the analyzer is equipped with a spiral microfluidic sorter, a filter, or tangential flow filtration for real-time separation of cells from media, and an in-line, microfluidic acoustic or electrical lyser.

17. The CAPCAS of claim 11 , further comprising a plurality of multichannel optical sensing modules.

18. The CAPCAS of claim 17 , wherein a first one of the multichannel optical sensing modules is coupled between the input plate and the biodevice plate, and a second one of the multichannel optical sensing modules is coupled between the biodevice plate and the output plate for measuring PO 2 , PCO 2 , pH, and/or optical density (OD) of the media entering and leaving each biodevice, respectively.

19. The CAPCAS of claim 18 , wherein a multichannel optical sensing module is coupled between the at least one microformulator and the input plate for tracking an intentionally injected bubble for measurement of flow rate, or identifying when a reservoir is emptied.

20. The CAPCAS of claim 11 , wherein each biodevice comprises a lid structure for controlling operation of the biodevice, wherein the lid structure comprises a fluidic control layer that contains motors that drive the pumps and valves, and a lid beneath the fluidic control layer, wherein the lid supports vertical tubes that deliver and remove fluid from the well, with a long tube reaching nearly to the bottom of the well to allow the pump P5 to remove when desired some or nearly all of the media and cells in the well, a medium length tube being connected to the pump P4 to provide continuous removal of media from the biodevice and deliver it to the at least one multimode output valve V3, and a short tube being connected to the pumps

Assignees

• Univ Vanderbilt

Inventors

• Reiserer Ronald S
• Gerken Gregory B
• Schaffer David K
• Wikswo John P

Classifications

• C12M27/12

  Roller bottles; Roller tubes · CPC title

• C12M23/58

  Reaction vessels connected in series or in parallel (combinations of bioreactors with other apparatus, C12M43/00) · CPC title

• C12M35/02

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12M29/10 Primary

  Perfusion · CPC title

• C12M35/04

  Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli · CPC title