Flow through electroporation instrumentation

• US 10,435,713 B2
• Filed: 09/28/2018
• Issued: 10/08/2019
• Est. Priority Date: 09/30/2017
• Status: Active Grant

• Current Assignee: Inscripta, Inc.
• Original Assignee: Inscripta, Inc.
• Inventors: Bernate, Jorge, Masquelier, Don
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US16/147,120
• Publication Number:
  US 20190100720A1
• Time in Patent Office: 375 Days
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/81 for yeasts
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The present disclosure provides a flow-through electroporation device configured for use as a stand-alone module or as one module in an automated multi-module processing system.

Claims

1. A flow-through electroporation (FTEP) device for introducing an exogenous material into cells in a fluid, wherein the FTEP device has a top surface and a bottom surface, the FTEP device comprising:

a. an inlet reservoir, an inlet, and a first inlet channel for receiving a fluid comprising cells and exogenous material into the FTEP device, wherein the inlet reservoir originates from the top surface of the FTEP device;

b. an outlet reservoir, an outlet, and an outlet channel for removing a fluid comprising transformed cells and exogenous material from the FTEP device, wherein the outlet reservoir originates from the top surface of the FTEP device;

c. a flow channel intersecting and positioned between the first inlet channel and the outlet channel, wherein the flow channel decreases in width to a dimension no smaller than at least 2×

diameter of the cells being electroporated to form a constriction of the flow channel between the first inlet channel and the outlet channel;

d. a first electrode and a second electrode each positioned in an electrode channel, wherein the electrode channels originate from the top surface of the FTEP device, wherein the first electrode is positioned between the first inlet channel and the constriction and wherein the second electrode is positioned between the constriction and the outlet channel;

wherein the electrodes are in fluid and electrical communication with fluid in the flow channel but not directly in the path of the cells traveling through the flow channel; and

wherein the electrodes apply one or more electric pulses to the cells in the fluid as they pass through the flow channel, thereby introducing exogenous material into the cells in the fluid; and

e. a pressure source, a flow sensor measurement device and a processor, wherein the processor is programmed such that, the pressure source pushes flow of the cells and exogenous material between the inlet reservoir and the outlet reservoir, the flow sensor measurement device senses flow of the cells and exogenous material and, when the flow sensor measurement device detects an increase in air flow indicative of fluid being completely drained from the inlet reservoir, the flow sensor measurement device communicates with the processor to communicate with the pressure source to pull flow of the cells and exogenous material between the outlet reservoir and the inlet reservoir, thereby reversing the flow of the cells and exogenous material.

2. The FTEP device of claim 1, wherein the FTEP device comprises a second inlet and a second inlet channel and further comprises a reservoir connected to the second inlet for introducing exogenous material into the FTEP device.

3. The FTEP device of claim 2, wherein the second inlet and second inlet channel are located between the first inlet and first inlet channel and the first electrode.

4. The FTEP device of claim 2, wherein the second inlet and second inlet channel are located between the second electrode and the outlet channel.

5. The FTEP device of claim 1, wherein device is configured for use with bacterial, yeast and mammalian cells.

6. The FTEP device of claim 1, wherein the two electrodes are located from 1 mm to 10 mm from one another.

7. The FTEP device of claim 1, wherein the narrowest part of the channel width is from 10 μ

M to 5 mm.

8. The FTEP device of claim 1, wherein the device further comprises one or more filters between the first inlet channel and the outlet channel.

9. An automated multi-module cell processing system, comprising the FTEP device of claim 1.