# US 2020/0399578 A1 · Devices, Methods, and Systems for Electroporation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399578 A1 - Document ID: US20200399578A1 - Status: Published application - Kind: A1 - Application number: 16/876,718 - Filed: 2020-05-18 - Published: 2020-12-24 - Earliest claimed: 2019-05-16 - Assignees: CyteQuest, Inc. - Applicants: CyteQuest, Inc. - Inventors: Tom Corso; Harold G. Craighead - Web page: https://patentgrep.com/patent/US20200399578A1 ## CPC - C12M 35/02: {Electrical or electromagnetic means, e.g. for electroporation or for cell fusion} - C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)} - C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Disclosed are a system, device, and method for the electroporation of cells. Systems, devices and methods for electroporation of living cells and the introduction of selected molecules into the cells utilizes a fluidic system where living cells and biologically active molecules flow through a channel that exposes them to electric fields, causing the molecules to be transferred across the cell membrane. The methods are particularly well suited for the introduction of DNA, RNA, drug compounds, and other biologically active molecules into living cells for use in cell-based therapies. ## Claims ### Claim 1 An electroporation device comprising a first-side support having at least one first-side electrode disposed at a region of an inner surface of the first-side support; a second-side support having at least one second-side electrode disposed at a region of an inner surface of the second-side support, wherein the at least one second-side electrode is a counterpart electrode of the at least one first-side electrode; and a fluid channel, having at least a portion of it between the first-side support and the second-side support, and having at least one fluid input and at least one fluid output, wherein the fluid channel allows fluid to flow continuously in at least one fluid stream toward the at least one fluid output, wherein the at least one first-side electrode and the at least one second-side electrode are positioned in the electroporation device to allow modulating the electric field as a predetermined function of time and position in at least one part of the fluid channel between the first-side support and the second-side support. ### Claim 2 (depends on 1) The electroporation device of claim 1, further comprising at least one voltage supplier, wherein the at least one first-side electrode and said counterpart electrode of the at least one first-side electrode are connected to said voltage supplier independently from any other electrodes of the electroporation device, and wherein said voltage supplier allows modulating the electric field as a function of time in said at least one part of the fluid channel. ### Claim 3 (depends on 2) The electroporation device of claim 2, comprising a plurality of first-side electrodes comprising the at least one first-side electrode, and comprising a plurality of second-side electrodes comprising the at least one second-side electrode, wherein at least two sets of counterpart electrodes operate independently from each other. ### Claim 4 (depends on 3) The electroporation device of claim 3, wherein at least three sets of counterpart electrodes operate independently from each other. ### Claim 5 (depends on 3) The electroporation device of claim 3, further comprising one or more additional voltage suppliers, wherein each voltage supplier is connected to a different set of counterpart electrodes in the electroporation device. ### Claim 6 (depends on 5) The electroporation device of claim 5, wherein said voltage suppliers allow forming an electric field as a function of time and position within the fluid channel that maximizes an outcome function that positively correlates with cell transfection efficiency and negatively correlates with cell mortality. ### Claim 7 (depends on 5) The electroporation device of claim 5, wherein said voltage suppliers allow forming an electric field as a function of time and position within the fluid channel that maximizes an outcome function that positively correlates with electrode durability. ### Claim 8 (depends on 5) The electroporation device of claim 5, wherein said voltage suppliers allow independently controlling any two or more of the following: (a) opening pores in cells in a fluid in the fluid channel; (b) driving molecules into cells in a fluid in the fluid channel; (c) measuring an electrical property of a fluid in the fluid channel; (d) concentrating molecules at a part of the fluid channel; (e) moving cells in a fluid in the fluid channel to a part of the fluid channel; and (f) rotating cells in a fluid in the fluid channel. ### Claim 9 (depends on 2) The electroporation device of claim 2, wherein said voltage supplier provides a voltage that has a periodic waveform. ### Claim at position 10 10-21. (canceled) ### Claim 22 An electroporation device comprising a first-side support having at least one first-side electrode disposed at a region of an inner surface of the first-side support; a second-side support having at least one second-side electrode disposed at a region of an inner surface of the second-side support, wherein the at least one second-side electrode is a counterpart electrode of the at least one first-side electrode; and a fluid channel, having at least a portion of it between the first-side support and the second-side support, and having at least two fluid inputs and at least one fluid output, wherein the fluid channel allows fluid to flow continuously in at least two fluid streams toward the at least one fluid output, wherein the fluid channel allows modulating the flow rate, chemical composition, or both the flow rate and chemical composition in the at least two fluid streams as a predetermined function of time, position, or both time and position. ### Claim 23 (depends on 22) The electroporation device of claim 22, further comprising at least one fluid supplier, wherein said fluid supplier allows modulating the flow rate and chemical composition in one of the at least two streams as a predetermined function of time, position, or both time and position independently from any other fluid stream within the fluid channel. ### Claim 24 (depends on 23) The electroporation device of claim 23, further comprising one or more additional fluid suppliers, wherein each fluid supplier is connected to a different fluid input in the fluid channel. ### Claim 25 (depends on 24) The electroporation device of claim 24, wherein said fluid suppliers allow modulating the flow rate and chemical composition in separate fluid streams as a function of time and position within the fluid channel to maximize the time a cell in a fluid stream is in its optimal medium. ### Claim 26 (depends on 24) The electroporation device of claim 24, wherein said fluid suppliers allow modulating the flow rate and chemical composition in separate fluid streams as a function of time and position within the fluid channel to minimize the time a cell in a fluid stream is in an electroporation medium. ### Claim 27 (depends on 22) The electroporation device of claim 22, wherein the fluid channel has at least three fluid inputs allowing fluid to flow continuously in at least three fluid streams toward the at least one fluid output. ### Claim at position 17 28-31. (canceled) ### Claim 32 A system, comprising an electroporation device, comprising a first-side support having at least one first-side electrode disposed at a region of an inner surface of the first-side support; a second-side support having at least one second-side electrode disposed at a region of an inner surface of the second-side support, wherein the at least one second-side electrode is a counterpart electrode of the at least one first-side electrode; and a fluid channel, having at least a portion of it between the first-side support and the second-side support, and having at least one fluid input and at least one fluid output, wherein the fluid channel allows fluid to flow continuously in at least one fluid stream toward the at least one fluid output, wherein the at least one first-side electrode and the at least one second-side electrode are positioned in the electroporation device to allow modulating the electric field as a predetermined function of time and position in at least one part of the fluid channel between the first-side support and the second-side support; and a fluid-delivery apparatus coupled to the electroporation device. ### Claim at position 19 33-34. (canceled) ### Claim 35 (depends on 32) The system of claim 32, further comprising a fluid interface that couples the fluid-delivery apparatus to the electroporation device. ### Claim at position 21 36-37. (canceled) ### Claim 38 (depends on 32) The system of claim 32, further comprising a cell processing module coupled to said electroporation device. ### Claim 39 (depends on 38) The system of claim 38, wherein said cell processing module is upstream from said electroporation device. ### Claim at position 24 40-42. (canceled) ### Claim 43 (depends on 39) The system of claim 39, further comprising an apheresis bag upstream of the cell processing module. ### Claim at position 26 44-57. (canceled)