WO2024107865A1

WO2024107865A1 - Arbitrary waveform electroporator circuit and device

Info

Publication number: WO2024107865A1
Application number: PCT/US2023/079856
Authority: WO
Inventors: Ivan RAZINKOV; Edwin CHIU
Original assignee: Synthego Corporation
Priority date: 2022-11-18
Filing date: 2023-11-15
Publication date: 2024-05-23

Abstract

An electroporator circuit and electroporator device can be used to deliver an electroporation signal to an electroporation load. The electroporator circuit can include an energy source, a filter, a converter, and a controller. The output of the electroporator circuit can be controlled based on an arbitrary waveform provided as input to the controller.

Description

ARBITRARY WAVEFORM ELECTROPORATOR CIRCUIT AND DEVICE

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority under 35 U.S.C. §119(e), to U.S. Provisional Application No. 63/426,475, as filed November 18, 2022, the contents of which are incorporated herein by reference in its entirety.

BACKGROUND

[0002] In microbiology, electroporation (sometimes referred to as electropermeabilization) can be used increase the permeability of cell membranes by applying an electrical field to cells. As a result, drugs, chemicals, foreign genes, electrodes, deoxyribonucleic acid (DNA), and other objects and substances can be introduced into cells (electrotransfer). During electroporation, an electric field can be passed across cells suspended in an electroporation cuvette to increase the permeability of the cell membranes. Electroporation can be used for gene therapy and other cellbased therapy, for example. More dynamic and configurable circuits and devices used for electroporation are generally desired for various purposes.

SUMMARY

[0003] Various implementations of the disclosure provide an electroporator circuit including an energy source, a filter, a converter, and a controller. The filter includes an inductor, a capacitor, and a filter output connectable to an electroporation load. The converter is connected to an output of the energy source and to an input of the filter. The controller is connected to the converter and is configured to receive an input comprising an arbitrary waveform and deliver a pulse-width- modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

[0004] Various implementations of the disclosure further provide a method for generating an electroporation waveform. The method includes providing a converter; providing a filter including an inductor, a capacitor, and a filter output connectable to an electroporation load; providing an energy source connected to the filter by the converter, where the converter is connected to an output of the energy source and an input of the filter; receiving, by a controller connected to the converter, an input including an arbitrary waveform; and delivering, using the controller, a pulse- width-modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

[0005] Various implementations of the disclosure further provide an electroporator device including an enclosure and an electroporator circuit. The electroporator circuit includes an energy source, a filter, a converter, and a controller. The filter includes an inductor, a capacitor, and a filter output connectable to an electroporation load. The converter is connected to an output of the energy source and to an input of the filter. The controller is connected to the converter and is configured to receive an input comprising an arbitrary waveform and deliver a pulse-width- modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The following drawings are provided to help illustrate various features of the disclosure and are not intended to limit the scope of the disclosure or exclude alternative configurations.

[0007] FIG. 1 shows an example electroporator circuit that can be used in an electroporator device.

[0008] FIGS. 2A-2B show example illustrations of an electroporator device that can include the electroporator circuit of FIG. 1.

[0009] FIG. 3 shows a block diagram illustrating example components of the electroporator circuit of FIG. 1.

[0010] FIG. 4 shows another block diagram illustrating example components of the electroporator circuit of FIG. 1.

[0011] FIG. 5 shows example illustrations of different states of the electroporator circuit of FIG. 1.

[0012] FIG. 6 shows example graphs of different signals associated with operation of the electroporator circuit of FIG. 1 over time.

[0013] FIGS. 7A-7D show an example schematic for the electroporator circuit of FIG. 1. [0014] FIG. 8 shows an example schematic for power supply components of the electroporator circuit of FIG. 1.

[0015] FIGS. 9A-9D show an example schematic for a controller of the electroporator circuit of FIG. 1 and associated connections.

[0016] FIG. 10 shows an example schematic for a capacitor charger of the electroporator circuit of FIG. 1.

[0017] FIGS. 11A-1 IB show an example schematic for a capacitor bank of the electroporator circuit of FIG. 1.

[0018] FIGS. 12A-12D show an example schematic for an amplifier of the electroporator circuit of FIG. 1.

[0019] FIGS. 13A-13C show an example schematic for another amplifier of the electroporator circuit of FIG. 1.

[0020] FIG. 14 shows an example schematic for yet another amplifier of the electroporator circuit of FIG. 1.

[0021] FIG. 15 shows an example schematic for parallel control connections of the electroporator circuit of FIG. 1

[0022] FIG. 16 shows a flow diagram illustration an example process for electroporation that can be implemented using the electroporator circuit of FIG. 1.

DETAILED DESCRIPTION

[0023] The foregoing and other aspects and advantages of the present disclosure will appear from the following description. In the description, reference is made to the accompanying drawings that form a part hereof, and in which there is shown by way of illustration one or more exemplary versions. These versions do not necessarily represent the full scope of the disclosure.

[0024] An electroporator device can use electricity to make cell membranes more permeable, for a variety of purposes. The electroporator device can include one or more electroporator circuits for generating waveforms used in the electroporation process. Some electroporator devices have one or more switched energy sources (e.g., capacitor banks) that are charged to a specific voltage and then connected to an electroporation load. However, these el ectr oporator devices can limit each electroporation waveform to a series of exponential decays starting at predetermined voltages, with the decay rate tied to the capacitance of the energy source(s) and the impedance of the electroporation load. Pulse shapes such as multi-step square profdes can be approximated using multiple capacitor banks charged to different voltages and switched to the load sequentially. However, in this case the number of steps is limited by the number of capacitor banks, and each segment of the electroporation waveform can only take one shape, namely an exponential decay tied to the impedance of the electroporation load. Some electroporator devices may also hide parameters from the end user, and only give out alphanumeric codes, limiting the ability of the user to customize the electroporation waveform.

[0025] Accordingly, disclosed herein are embodiments of an electroporator device and associated electroporator circuit which can be designed to deliver an arbitrary, user-defined waveform shape to an electroporation load (sample) that is independent of the capacitance of the energy source, the impedance of the electroporation load, and the number of capacitor banks used. Various embodiments of the disclosed electroporator device may be used with various systems including automated cell processing systems such as those disclosed in International/PCT Application PCT/US2022/012820, published as WO 2022/155610, which is incorporated herein by reference in its entirety.

[0026] In this design, the number of waveform setpoints is not limited by the number of capacitor banks, and the electroporator circuit and device can provide arbitrary waveforms with as few as a single capacitor bank, thereby reducing system complexity. Also, in this design, the waveform is not limited to exponential decays, but rather can take any shape as defined by the user of the electroporator. An electroporator device and associated electroporator circuit can also be designed to provide a dual-pulse waveform, such as a dual-pulse of 200 volts for 1 millisecond, or 116 volts for 30 milliseconds. The duration of each pulse can be controlled programmatically, and the voltage value can be controlled mechanically via a dial, in some implementations. The output waveform produced using the electroporation device and associated electroporation circuit can provide improved transfection conditions for T cells, Jurkat cells, Hematopoietic stem cells (HSC), and monocytes, among other types of cells. The electroporator device and associated electroporator circuit can help users explore and understand the electroporation conditions that lead to improved transfection, as well as knock-out and knock-in efficiencies, for example. [0027] FIG. 1 shows an example electroporator circuit 100 that can be used in an el ectr oporator device. Electroporator circuit 100, as shown in the example illustration of FIG. 1, includes an energy source 110, a filter 120, a converter 130, and a controller 140. Electroporator circuit 100 can be used to deliver arbitrary waveforms consisting of an arbitrary number of time and voltage setpoints. Electroporator circuit 100 can also handle large electroporation loads (e.g., 1000 microliters (pL) or more). Electroporator circuit 100 can include converter 130 placed between energy source 110 and an electroporation load 150 (discussed with respect to FIG. 3 and FIG. 4), instead of a switch. Electroporator circuit 100 can use feedforward control, rather than feedback control, to compensate for the changing source voltage of energy source 110 and ensure dynamic stability and a fast response time. In some implementations, electroporator circuit 100 can be provided on a single printed circuit board (PCB), however other implementations are possible and contemplated.

[0028] Energy source 110 is shown as a capacitor bank, which in this particular embodiment includes ten individual capacitors in which two strings of five capacitors that are connected in series are in turn connected in parallel. The capacitors used to implement this example capacitor bank can be of the same or similar electrical ratings. It is important to note that energy source 110 can be implemented using a variety of different configurations of capacitors, including using only a single capacitor, among other possible implementations that can include electrical components in addition to and/or in place of capacitors. Energy source 110 can generally be used to store energy within electroporator circuit 100 that can ultimately be used to deliver an electroporation signal to an electroporation load. Energy source 110 can include and/or be connected to a sensor 112, a charger 116, and a safety mechanism 170, as discussed in more detail below, for example, among other components. In some implementations, energy source 110 can be designed to provide high power (e.g., up to 300 kilowatts) within electroporator circuit 100. The capacitors in the capacitor bank can discharge upon closing of converter 130, and accordingly monitoring the voltage of energy source 110 using sensor 112 and appropriately charging energy source 110 using charger 116 can be important.

[0029] Converter 130 can be used to modulate the voltage delivered from energy source 110 to electroporation load 150 in response to a command received from a user (e.g., arbitrary waveform 142 input by user device 400, as discussed in more detail below). Converter 130 can also be used to compensate for voltage changes in energy source 110, such as the exponential voltage decay in a capacitor bank. Converter 130 can be implemented as a buck converter, for example, using a high-power metal-oxide-semiconductor field-effect transistor (MOSFET) or an insulated-gate bipolar transistor (IGBT). Converter 130 can switch energy source 110 on and off at a rate higher than the desired resolution of the arbitrary waveform. For example, if the desired resolution is 10 kilohertz (kHz), converter 130 can implement switching at a higher frequency such as 20 kHz to 50 kHz, among other possible examples. The switching performed by converter 130 can result in a constant frequency pulse-width-modulated (PWM) output with a controllable duty cycle. The pulse-width-modulated output can be a discontinuous current with an average voltage of the voltage of energy source 110 multiplied by the duty cycle.

[0030] Converter 130 can convert energy from energy source 110 at one voltage to a commanded target voltage at high speed. Linear (variable resistance) amplifiers may be difficult to use at high power applications because of the heat dissipation involved, where in some examples up to 50% of power is lost to heat in the semiconductor. However, switching amplifiers can be used to implement converter 130 using semiconductors that are either fully on (very low resistance) or fully off (very high resistance), and accordingly these semiconductors may only have to dissipate around 5% of power as heat, in some examples. By varying the duty cycle of the converter 130, the output voltage can be changed as a ratio of the input voltage. The ratio of output voltage to input voltage, accordingly, can be equal to the duty cycle.

[0031] Filter 120 can convert the discontinuous current (pulse-width-modulated output) generated by converter 130 into a continuous current to be fed into electroporation load 150. Filter 120 can be implemented using an inductor and a capacitor (e.g., inductor 122 and capacitor 124 discussed below) to form an LC filter. The inductor can store energy during the on state of the switching cycle implemented by converter 130 and release the energy during the off-state of the switching cycle to a freewheeling diode (e.g., freewheeling diode 160 discussed below) that carries the current during the off-state. Freewheeling diode 160, in some examples, can be separated from capacitor 124 by converter 130 such that in an off-state of converter 130 freewheeling diode 160 is connected in series with electroporation load 150 and inductor 122. The output capacitor can then smooth the current so that the final output of electroporator circuit 100 applied to electroporation load 150 can be approximated as a constant voltage equal to the average of the pulse-width-modulated output of converter 130. The inductor of filter 120 can convert an on-off input into a continuously variable output by forcing the output current to take on a rate-limited current ramp instead of an instantaneous rise. Due to this ramp, a variable amount of time can become a variable amount of current, and thus a variable amount of voltage into a constant impedance. The capacitor of filter 120 can smooth the output waveform (e.g., a sawtooth wave).

[0032] Controller 140 can be implemented as a microcontroller that commands the duty cycle of converter 130 according to a desired waveform (e.g., arbitrary waveform 142 discussed below) and a sensed voltage associated with energy source 110. Controller 140 can use feedforward control to compensate for the changing source voltage of energy source 110 and ensure dynamic stability and a fast response time without saucing potential instability in certain load conditions. In simple form, the general formula for control implemented by controller 140 can be expressed as: Duty Cycle = ^{Command v}°^lta9^e Controller 140 can be implemented using a variety of

different types and configurations of controllers, including using more than one microcontroller, single or multiple core processors, and other types of processing circuitry and hardware components depending on the application. For example, controller 140 can include different quantities and configurations of input/output (I/O) terminals, analog-to-digital converters (ADC) and/or digital-to-analog converters (DAC), and clocking mechanisms.

[0033] Various design parameters of electroporator circuit 100 can be modified to adjust the capabilities and limitations of electroporator circuit 100. In some implementations, electroporator circuit 100 can be designed to provide a maximum output voltage of 450 volts and a minimum output voltage of 0 volts. To allow for a maximum electroporation load of 1000 microliters with a 450-volt pulse delivered (about 4 ohms of impedance), electroporator circuit 100 can be designed to provide a maximum output current of 120 amperes. In some implementations, electroporator circuit 100 can be designed to have a maximum slew rate of 6 amperes per microsecond, a maximum total waveform energy of 840 joules (safety limit to prevent overheating of electroporation load and components of electroporator circuit 100), a maximum waveform time resolution of 20 kilohertz, and a delivered waveform feedback of 20 kilohertz. Also, communications between controller 140 and associated components can occur via serial communications over universal serial bus (USB) universal asynchronous receiver-transmitter (UART) at a 115200 baud rate. However, it is important to note these values are provided as examples to help illustrate the functionality of electroporator circuit 100, and the skilled person will understand various design parameters of electroporator circuit 100 can be changed to alter these values.

[0034] FIGS. 2A-2B show example illustrations of an electroporator device 200 that can include electroporator circuit 100. As shown, electroporator device 200 includes an enclosure 210 that houses electroporator circuit 100 and other electrical and mechanical components. Electroporator device 200 can include one or more fans for circulating air within enclosure 210 to provide cooling for electrical components that can generate excessive heat during the electroporation process. In some cases, the electroporator device 200 can pass thousands of volts across electroporation load 150 during the electroporation process, and accordingly dissipation of heat within enclosure 210 can be of high importance for safety reasons. Electroporator device 200 can also include a safety shutdown button or switch, as well as various electrical connection and wiring interface for connecting electroporator device 200 to electroporation load 150 and a power supply, for example. FIG. 2A specifically shows electroporator device 200, where enclosure 210 is not transparent such that the interior of electroporator device 200 cannot be seen. FIG. 2B specifically shows electroporator device 200, where enclosure 210 is transparent such that the interior of electroporator device 200 and electroporator circuit 100 can be seen. It is important to note that a variety of different implementations of an electroporator device such as electroporator device 200 can be designed and manufactured using electroporator circuit 100.

[0035] FIG. 3 shows a block diagram illustrating example components of electroporator circuit 100. FIG. 3 illustrates relationships between energy source 110, filter 120, converter 130, controller 140, and electroporation load 150. FIG. 2 also shows a sensor 112 that can measure one or more variables associated with energy source 110, a sensor output 114 that can be generated by sensor 112 and received as input by controller 140, an arbitrary waveform 142 that can be received as input by controller 140, and a pulse-width-modulated (PWM) control signal 144 output from controller 140 and provided as input to converter 130. FIG. 3 also illustrates fdter 120, including inductor 122 and capacitor 124, and a filter output 126 that can be generated by filter 120 and provided to electroporation load 150. Filter output 126 can thus be an electroporation waveform provided to electroporation load 150. FIG. 3 illustrates a control path including the flow of arbitrary waveform 142 into controller 140, and the flow of pulse-width-modulated control signal 144 from controller 140 into converter 130, as well as a sensing path including the flow of sensor output 114 from sensor 112 to controller 140. FIG. 3 also illustrates an energy path including the flow of energy from energy source 1 10, through converter 130 and filter 120, and to electroporation load 150. The block diagram illustrated in FIG. 3 shows representation of electroporator circuit 100 that is provided to help illustrate the functionality of el ectr oporator circuit 100.

[0036] Energy source 110 can be implemented as a capacitor bank, including a plurality of individual capacitor components as illustrated for example in FIG. 1. Energy source 110 can generally be used to store energy within electroporator circuit 100 that can ultimately be used to deliver an electroporation signal to electroporation load 150. In some implementations, energy source 110 can be designed to provide high power (e g., up to 40 kilowatts) within electroporator circuit 100. Energy source 110 can discharge upon closing of converter 130, and this discharging can be monitored by sensor 112. Sensor 112 can be designed sense one or more voltages associated with energy source 110, such as voltages across one or more capacitors and/or strings of connected capacitors, and generate sensor output 114 based on these sensed voltages. Sensor 112 can also be designed to sense parameters associated with energy source 110 such as current levels, impedance, leakage, temperature, and other parameters associated with energy source 110. Sensor 112 can be implemented as one individual sensor components, or using multiple separate sensor components. Sensor 112 can provide sensor output 114 to controller 140 such that controller 140 can affect operation of electroporator circuit 100 based on sensor output 114. In some implementations, sensor 112 can also sense various parameters associated with charger 116, as discussed in more detail below.

[0037] Electroporation load 150 can be implemented in a variety of ways, including using a variety of different load sizes and types of cells for electroporation. For example, electroporation load can be a large load of around 1 milliliter, 5 milliliters, 10 milliliters, or more, or a smaller load such as 50 microliters. Electroporation load 150 can include different types of cells such as T cells, Jurkat cells, Hematopoietic stem cells, and monocytes, among other types of cells. Electroporation load 150 can be provided in the form of an electroporation cuvette, made of a transparent material, and including a liquid containing the cells. Filter output 126 can be provided to electroporation load 150 by electroporator circuit 100 to increase the permeability of the cell membranes of the cells suspended in the electroporation cuvette. As a result, drugs, chemicals, foreign genes, electrodes, DNA, and other objects and substances can be introduced into the cells. [0038] FIG. 4 shows another block diagram illustrating example components of electroporator circuit 100. The block diagram shown in FIG. 4 is similar to the block diagram shown in FIG. 3, however the block diagram in FIG. 4 shows a bit of a different perspective and some additional components and signals associated with electroporator circuit 100. For example, a user device 400 is shown in FIG. 4 that can be used to provide arbitrary waveform as input to controller 140. FIG. 4 also shows a safety mechanism 170 that can be implemented within electroporator circuit 100 to help prevent overheating and other potential hazards that may arise during operation of electroporator circuit 100. FIG. 4 also shows a charger connected to controller 140 and energy source 110 that can be used to charge energy source 110. FIG. 4 also shows an output sensor 128 that can generate a signal indicative of the actual waveform provided to electroporation load 150 and provide that signal back to controller 140. Controller 140 can then provide the signal indicative of the actual waveform provided to electroporation load 150 to user device 400 for verification. FIG. 4 also shows different example components of controller 130, including analog- to-digital converter terminals, general purpose input/output terminals, clocked input/output terminals. FIG. 4 also shows an example gate drive interlock signal provided to converter 130 by controller 140.

[0039] Charger 116 can be implemented as a flyback converter, for example, that takes a 24- volt input and steps it up to 250 volts. Charger 116 can be a buck-boost converter with an inductor split to form a transformer, with the step up in voltage occurs with galvanic isolation of the input and output voltages. Charger 116 can also include a rectifying diode, in some implementations. Charger 116 can also include one or more switches to turn the charger on and off. During an on state of charger 116, energy can be transferred from the input voltage source (e.g., voltage provided by controller 120) to the transformer of charger 116. During an off state of charger 116, energy can be transferred from the transformer of charger 116 to the output load (e.g., one or more capacitors of energy source 110). Controller 140 can control operation of charger 116, including for example by providing an input voltage to charger 116 and/or operating one or more switches of charger 116 to move charger 116 between the on state and the off state, based on output 114 of sensor 112. Various parameters of charger 116 can be monitored by controller 140, and controller 140 can terminate charging of energy source 1 10 using charger 116 when a software-controlled target threshold voltage level is reached, for example. Controller 140 and charger 116 can provide some level of hysteresis (e g., gap between turn-on and turn-off trigger levels) to avoid chattering effects that may occur.

[0040] Safety mechanism 170 can be used to remove power from electroporator device 200 under potentially hazardous conditions. A relay can be provided within enclosure 210 that allows a low-current 24-volt interlock loop to control power to electroporator device 200, for example. This circuit can then be run to any number of emergency switches, sensors, programmable logic controllers (PLCs), interlock switches, and other devices and components, either internal or external to enclosure 210. Any break in the low-current loop can then cut off power to electroporator device 200. Moreover, the switches in electroporator device 200, and electroporator circuit 100, can be positive controlled. Thereby, if power is removed, the switches can default to an off condition, and all outputs can be disconnected from energy source 100. Further, an onboard safety relay can be provided and held open by the input power. Once power is removed, this relay can close under spring force and discharge energy source 110 through resistors, for example. Electroporator circuit 100 can also include one or more labeled light-emitting diodes (LEDs) to indicate the presence of one or more hazardous voltages on-board, for the safety of anyone servicing the board. Moreover, as shown in FIG. 4, a two-stage control can be provided on the gate driver of converter 130, where controller 140 first can arm the gate driver, and then provide the command signal, to turn converter 130 on. This can guard against unintentional firing of converter 130. Each of these described components and functionality can be provided as part of safety mechanism 170 for protection during operation of electroporator circuit 100 and electroporator device 200.

[0041] To confirm the actual waveform delivered by electroporation circuit 100 to electroporation load 150, controller 140 can receive as input voltage and current measurements taken from filter output 126 by output sensor 128 (e.g., at ADC inputs of controller 140). This actual delivered waveform can then be recorded by controller 140 and provided back to the user via user device 400. This verification functionality can be useful because of the rate-limited nature of converter 130, the actual delivered waveform can have a frequency -limited shape. Especially for very sharp waveforms, the rate-limited nature of converter 130 can significantly affect the actual delivered waveform, which can make the ability to provide the actual output waveform to the user for verification in this manner valuable. [0042] User device 400 can be implemented in a variety of different ways but can generally receive inputs from a user and provide outputs to a user, as well as communicate with controller 140 using a variety of different protocols. User device 400 can include components such as a processor, memory, a display, inputs (e.g., a keyboard, a mouse, a graphical user interface, a touchscreen display, one or more actuatable buttons, etc.), communication devices (e.g., an antenna and appropriate corresponding circuitry), etc. User device 400 can be implemented as a mobile phone (e.g., a smart phone), a personal digital assistant (PDA), a laptop, a notebook, a netbook computer, a tablet computing device, and other similar types of electronic devices, for example. Through user device 400, a user of el ectr oporator circuit 100 and el ectr oporator device 200 can view the actual waveform delivered by electroporation circuit 100 to electroporation load 150, for example as sensed by output sensor 128. Through user device 400, a user of electroporator circuit 100 and electroporator device 200 can also provide arbitrary waveform 142 as input to controller 140, such that controller 140 affects operation of converter 130 in accordance with arbitrary waveform 142, for example.

[0043] FIG. 5 shows example illustrations of both an on state 510 and an off state 520 of electroporator circuit 100. In FIG. 5, freewheeling diode 160 (sometimes referred to as a flyback diode, snubber diode, suppressor diode, etc.) is shown to be connected in parallel with energy source 110, capacitor 124, and electroporation load 150. Also, converter 130 is shown to be connected in series with inductor 122. In on state 510, as shown, converter 130 is switched closed, and energy flows from energy source 110 through electroporation circuit 100. In the on state, freewheeling diode 160 is inactive. In off state 520, as shown, converter 130 is switched open, and energy source 110 is disconnected from the other components of electroporator circuit 100. Also, in off state 520, freewheeling diode 160 is active to prevent to prevent any sudden voltage spikes seen across other components of electroporation circuit 100 that may occur when energy source 110 is cut off due to the opening of converter 130. Freewheeling diode 160 can allow current to continue to circulate within electroporator circuit 100 even when converter 130 is switched off, and electroporator circuit 100 is in off state 520. Freewheeling diode 160 can generally be connected in series with electroporation load 150 and inductor 122.

[0044] FIG. 6 shows example graphs of different signals associated with operation of electroporator circuit 100 overtime. Graph 610 shows the status of converter 130 overtime, where converter 130 is either switched on or switched off. Graph 620 shows the current flowing through inductor 122 over time. As shown in FIG. 6, the slope of the increasing current through inductor 122 when converter 130 is switched on can be expressed using the equation Slope = input Voltage — Output Voltage

slope of the decreasing current through inductor 122 when

Inductance

. . . . . . . . - Output Voltage > . converter 130 is switched on can be expressed using the equation Slope = — _Inductance — ■ fj^raph

630 shows the current flowing through freewheeling diode 160 over time. As shown, no current flows through freewheeling diode 160 when converter 130 is switched on, but a spike in current flowing through freewheeling diode 160 occurs when converter 130 is switched off, and this current flowing through freewheeling diode 160 linearly decreases with time following the initial spike. Graph 640 shows the input current provided by energy source 110 over time, where a spike in current provided by energy source 110 occurs when converter 130 is switched on, and this current provided by energy source 110 linearly increases with time following the initial spike. However, when converter 130 is switched off, the current flow from energy source 110 is cut off and goes to zero. These graphs shown in FIG. 6 are provided as examples to help illustrate the functionality of electroporator circuit 100, and the skilled person will understand various design parameters of electroporator circuit 100 can be changed to alter the functionality of electroporator circuit 100 such that it varies from what is shown in the graphs of FIG. 6.

[0045] FIGS. 7A-7D show an example schematic for electroporator circuit 100, including components such as energy source 110, charger 116, and controller 140. FIG. 8 shows an example schematic for power supply components of electroporator circuit 100. FIGS. 9A-9D show an example schematic for controller 140 of electroporator circuit 100 and associated connections. FIG. 10 shows an example schematic for charger 116 of electroporator circuit 100. FIGS. 11A- 1 IB show an example schematic for energy source 110 of electroporator circuit 100. FIGS. 12A- 12D show an example schematic for an amplifier of electroporator circuit 100. FIGS. 13A-13C show an example schematic for another amplifier of electroporator circuit 100. FIG. 14 shows an example schematic for yet another amplifier of electroporator circuit 100. FIG. 15 shows an example schematic for parallel control connections of electroporator circuit 100. It will be appreciated that the schematics shown in FIGS. 7A-15 are provided to help illustrate an example of how electroporator circuit 100 can be implemented. The skilled person will understand that modifications to these schematics are possible and contemplated, to adjust various operational parameters and functionality of electroporator circuit 100 without departing from the scope of the present disclosure.

[0046] FIG. 16 shows a flow diagram illustration an example process 1600 for electroporation that can be implemented using the electroporator circuit of FIG. 1. Process 1600 can include using electroporator circuit 100 and electroporator device 200 to provide more dynamic and configurable electroporation signals to an electroporation load. Process 1600 can also be used to handle electroporation loads of large sizes, for example electroporation loads of around 1000 microliters. Process 1600 can be used to deliver an electroporation signal to an electroporation load that is an electroporation cuvette filled with cells such as T cells, Jurkat cells, Hematopoietic stem cells, and monocytes, among other types of cells, for example. Process 1600 can be used to deliver an arbitrary, user-defined waveform shape to an electroporation load that is independent of the capacitance of the energy source, the impedance of the electroporation load, and the number of capacitors used as an energy source, for example.

[0047] At step 1610, an energy source that is coupled to a filter by a converter is provided. For example, energy source 110 connected to filter 120 by converter 130 can be provided as part of electroporation circuit 100. Energy source 110 can be a capacitor bank, including multiple individual capacitors connected together in various configurations depending on the application. Converter 130 can be implemented as a MOSFET or an IGBT, and can be controlled by controller 140 and used to modulate the voltage delivered form energy source 110 to filer 120, and then ultimately to electroporation load 150. Filter 120 can include inductor 122, capacitor 124, and freewheeling diode 160. Filter output 120 can be connectable to an electroporation load of at least 1000 microliters. Sensor 112 can sense one or more parameters, including one or more voltages, associated with energy source 110 and provide sensor output 114 to controller 140.

[0048] At step 1620, an input is received including an arbitrary waveform. For example, controller 140 can receive arbitrary waveform 142 as input from user device 400. The arbitrary waveform can include a variety of different waveform parameters, including frequencies of up to 10 kilohertz, in some examples. Through user device 400, a user of electroporator circuit 100 and electroporator device 200 can view the actual waveform delivered by electroporation circuit 100 to electroporation load 150, for example as sensed by output sensor 128, in addition to providing arbitrary waveform 142 as input to controller 140. [0049] At step 1630, a pulse-width-modulated control signal is delivered to the converter based on the arbitrary waveform. For example, controller 140 can deliver pulse-width-modulated control signal 144 to converter 130 based on arbitrary waveform 142 received as input by controller 140 from user device 400. Accordingly, electroporation circuit can control the actual waveform provided to electroporation load 150 based on arbitrary waveform 142. Energy can be passed form energy source 110 to filter 120 based on arbitrary waveform 142, and the user can verify the actual waveform provided to electroporation load 150 by viewing the data generated by output sensor 128. Accordingly, process 1600 can be used to deliver an arbitrary waveform to electroporation load 150.

[0050] The present disclosure has described one or more preferred claims, and it should be appreciated that many equivalents, alternatives, variations, and modifications, aside from those expressly stated, are possible and within the scope of the invention.

[0051] It is to be understood that the disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the following drawings. The disclosure is capable of other claims and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” or “having” and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless specified or limited otherwise, the terms “mounted,” “connected,” “supported,” and “coupled” and variations thereof are used broadly and encompass both direct and indirect mountings, connections, supports, and couplings. Further, “connected” and “coupled” are not restricted to physical or mechanical connections or couplings.

[0052] As used herein, unless otherwise limited or defined, discussion of particular directions is provided by example only, with regard to particular claims or relevant illustrations. For example, discussion of “top,” “front,” or “back” features is generally intended as a description only of the orientation of such features relative to a reference frame of a particular example or illustration. Correspondingly, for example, a “top” feature may sometimes be disposed below a “bottom” feature (and so on), in some arrangements or claims. Further, references to particular rotational or other movements (e g., counterclockwise rotation) is generally intended as a description only of movement relative a reference frame of a particular example of illustration.

[0053] In some claims, aspects of the disclosure, including computerized configurations of methods according to the disclosure, can be implemented as a system, method, apparatus, or article of manufacture using standard programming or engineering techniques to produce software, firmware, hardware, or any combination thereof to control a processor device (e.g., a serial or parallel general purpose or specialized processor chip, a single- or multi-core chip, a microprocessor, a field programmable gate array, any variety of combinations of a control unit, arithmetic logic unit, and processor register, and so on), a computer (e.g., a processor device operatively coupled to a memory), or another electronically operated controller to implement aspects detailed herein. Accordingly, for example, claims of the disclosure can be implemented as a set of instructions, tangibly embodied on a non-transitory computer-readable media, such that a processor device can implement the instructions based upon reading the instructions from the computer-readable media. Some claims of the disclosure can include (or utilize) a control device such as an automation device, a special purpose or general-purpose computer including various computer hardware, software, firmware, and so on, consistent with the discussion below. As specific examples, a control device can include a processor, a microcontroller, a field- programmable gate array, a programmable logic controller, logic gates etc., and other typical components that are known in the art for configuration of appropriate functionality (e.g., memory, communication systems, power sources, user interfaces and other inputs, etc.).

[0054] The term “article of manufacture” as used herein is intended to encompass a computer program accessible from any computer-readable device, carrier (e.g., non-transitory signals), or media (e.g., non-transitory media). For example, computer-readable media can include but are not limited to magnetic storage devices (e.g., hard disk, floppy disk, magnetic strips, and so on), optical disks (e.g., compact disk (CD), digital versatile disk (DVD), and so on), smart cards, and flash memory devices (e.g., card, stick, and so on). Additionally it should be appreciated that a carrier wave can be employed to carry computer-readable electronic data such as those used in transmitting and receiving electronic mail or in accessing a network such as the Internet or a local area network (LAN). Those skilled in the art will recognize that many modifications may be made to these configurations without departing from the scope or spirit of the claimed subject matter. [0055] Certain operations of methods according to the disclosure, or of systems executing those methods, may be represented schematically in the figures or otherwise discussed herein. Unless otherwise specified or limited, representation in the figures of particular operations in particular spatial order may not necessarily require those operations to be executed in a particular sequence corresponding to the particular spatial order. Correspondingly, certain operations represented in the figures, or otherwise disclosed herein, can be executed in different orders than are expressly illustrated or described, as appropriate for particular claims of the disclosure. Further, in some claims, certain operations can be executed in parallel, including by dedicated parallel processing devices, or separate computing devices configured to interoperate as part of a large system.

[0056] As used herein in the context of computer configuration, unless otherwise specified or limited, the terms “component,” “system,” “module,” and the like are intended to encompass part or all of computer-related systems that include hardware, software, a combination of hardware and software, or software in execution. For example, a component may be, but is not limited to being, a processor device, a process being executed (or executable) by a processor device, an object, an executable, a thread of execution, a computer program, or a computer. By way of illustration, both an application running on a computer and the computer can be a component. One or more components (or system, module, and so on) may reside within a process or thread of execution, may be localized on one computer, may be distributed between two or more computers or other processor devices, or may be included within another component (or system, module, and so on).

[0057] In some configurations, devices or systems disclosed herein can be utilized or installed using methods embodying aspects of the disclosure. Correspondingly, description herein of particular features, capabilities, or intended purposes of a device or system is generally intended to inherently include disclosure of a method of using such features for the intended purposes, a method of implementing such capabilities, and a method of installing disclosed (or otherwise known) components to support these purposes or capabilities. Similarly, unless otherwise indicated or limited, discussion herein of any method of manufacturing or using a particular device or system, including installing the device or system, is intended to inherently include disclosure, as claims of the disclosure, of the utilized features and implemented capabilities of such device or system. [0058] As used herein, unless otherwise defined or limited, ordinal numbers are used herein for convenience of reference based generally on the order in which particular components are presented for the relevant part of the disclosure. In this regard, for example, designations such as “first,” “second,” etc., generally indicate only the order in which the relevant component is introduced for discussion and generally do not indicate or require a particular spatial arrangement, functional or structural primacy or order.

[0059] As used herein, unless otherwise defined or limited, directional terms are used for convenience of reference for discussion of particular figures or examples. For example, references to downward (or other) directions or top (or other) positions may be used to discuss aspects of a particular example or figure, but do not necessarily require similar orientation or geometry in all installations or configurations.

[0060] Also as used herein, unless otherwise limited or defined, “or” indicates a non-exclusive list of components or operations that can be present in any variety of combinations, rather than an exclusive list of components that can be present only as alternatives to each other. For example, a list of “A, B, or C” indicates options of: A; B; C; A and B; A and C; B and C; and A, B, and C. Correspondingly, the term “or” as used herein is intended to indicate exclusive alternatives only when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of” For example, a list of “one of A, B, or C” indicates options of: A, but not B and C; B, but not A and C; and C, but not A and B. A list preceded by “one or more” (and variations thereon) and including “or” to separate listed elements indicates options of one or more of any or all of the listed elements. For example, the phrases “one or more of A, B, or C” and “at least one of A, B, or C” indicate options of: one or more A; one or more B; one or more C; one or more A and one or more B; one or more B and one or more C; one or more A and one or more C; and one or more of A, one or more of B, and one or more of C. Similarly, a list preceded by “a plurality of’ (and variations thereon) and including “or” to separate listed elements indicates options of multiple instances of any or all of the listed elements. For example, the phrases “a plurality of A, B, or C” and “two or more of A, B, or C” indicate options of: A and B; B and C; A and C; and A, B, and C.

[0061] Furthermore, in those instances where a convention analogous to “at least one of A, B and C, etc.” is used, in general such a construction is intended in the sense of one having ordinary skill in the art would understand the convention (e.g., “a system having at least one of A, B and C” would include but not be limited to systems that have A alone, B alone, C alone, A and B together, A and C together, B and C together, and/or A, B, and C together.). 1 1 will be further understood by those within the art that virtually any disjunctive word and/or phrase presenting two or more alternative terms, whether in the description or figures, should be understood to contemplate the possibilities of including one of the terms, either of the terms, or both terms. For example, the phrase “A or B” will be understood to include the possibilities of “A” or ‘B or “A and B.”

[0062] This discussion is presented to enable a person skilled in the art to make and use claims of the disclosure. Various modifications to the illustrated examples will be readily apparent to those skilled in the art, and the generic principles herein can be applied to other examples and applications without departing from the principles disclosed herein. Thus, claims of the disclosure are not intended to be limited to claims shown, but are to be accorded the widest scope consistent with the principles and features disclosed herein and the claims below. The following detailed description is to be read with reference to the figures, in which like elements in different figures have like reference numerals. The figures, which are not necessarily to scale, depict selected examples and are not intended to limit the scope of the disclosure. Skilled artisans will recognize the examples provided herein have many useful alternatives and fall within the scope of the disclosure.

[0063] Various features and advantages of the disclosure are set forth in the following claims.

Claims

CLAIMS What is claimed is:

1. An electroporator circuit, comprising: an energy source; a filter comprising an inductor, a capacitor, and a filter output connectable to an electroporation load; a converter connected to an output of the energy source and to an input of the filter; and a controller connected to the converter and configured to: receive an input comprising an arbitrary waveform; and deliver a pulse-width-modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

2. The electroporator circuit of claim 1, wherein the energy source comprises a capacitor bank.

3. The electroporator circuit of claim 1 or claim 2, further comprising a sensor configured to provide a sensor output to the controller, the sensor output indicative of a voltage associated with the energy source.

4. The electroporator circuit of claim 3, wherein the controller is further configured to receive the sensor output and deliver the pulse-width-modulated control signal based on the sensor output.

5. The electroporator circuit of any one of the preceding claims, wherein converter comprises a metal-oxide-semiconductor field-effect transistor or an insulated-gate bipolar transistor.

6. The electroporator circuit of any one of the preceding claims, wherein the filter further comprises a freewheeling diode connected in series with the electroporation load and the inductor.

7. The electroporator circuit of any one of the preceding claims, wherein the filter output is connectable to an electroporation load of at least 1000 microliters.

8. The electroporator circuit of any one of the preceding claims, wherein the controller is configured to receive the arbitrary waveform as input from a user device, and wherein the arbitrary waveform comprises a frequency of up to 10 kilohertz and a resolution of up to 20 kilohertz.

9. A method for generating an electroporation waveform, comprising: providing a converter; providing a filter comprising an inductor, a capacitor, and a filter output connectable to an electroporation load; providing an energy source connected to the filter by the converter, wherein the converter is connected to an output of the energy source and an input of the filter; receiving, by a controller connected to the converter, an input comprising an arbitrary waveform; and delivering, using the controller, a pulse-width-modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

10. The method of claim 9, wherein providing the energy source comprises providing a capacitor bank.

11 . The method of claim 9 or claim 10, further comprising: providing a sensor configured to generate a sensor output indicative of a voltage associated with the energy source; and receiving, by the controller, the sensor output from the sensor

12. The method of claim 11, wherein delivering, using the controller, the pulse-width- modulated control signal to the converter comprises delivering, using the controller, the pulse- width-modulated control signal to the converter based on the sensor output.

13. The method of any one of claims 9-12, wherein providing the converter comprises providing a metal-oxide-semiconductor field-effect transistor or an insulated-gate bipolar transistor.

14. The method of any one of claims 9-13, wherein providing the filter further comprises providing the filter comprising a freewheeling diode connected in series with the electroporation load and the inductor.

15. The method of any one of claims 9-14, wherein the filter output is connectable to an electroporation load of at least 1000 microliters.

16. The method of any one of claims 9-15, wherein receiving, by the controller, the arbitrary waveform comprises receiving, by the controller, the arbitrary waveform as input from a user device.

17. An electroporator device, comprising: an enclosure; and an electroporator circuit disposed within the enclosure, the electroporator circuit comprising: an energy source; a filter comprising an inductor, a capacitor, and a filter output connectable to an electroporation load; a converter connected to an output of the energy source and to an input of the LC filter; and a controller connected to the converter and configured to: receive an input comprising an arbitrary waveform; and deliver a pulse-width-modulated control signal to the converter based on the arbitrary waveform such that the output of the filter is based on the arbitrary waveform.

18. The electroporator device of claim 17, wherein the energy source comprises a capacitor bank, and wherein the electroporator circuit further comprises a sensor configured to provide a sensor output to the controller, the sensor output indicative of a voltage associated with the energy source

19. The el ectr oporator device of claim 18, wherein the controller is further configured to receive the sensor output and deliver the pulse-width-modulated control signal based on the sensor output.

20. The electroporator device of claim 18, wherein the converter comprises a metal-oxide- semiconductor field-effect transistor or an insulated-gate bipolar transistor, and wherein the filter further comprises a freewheeling diode connected in series with the electroporation load and the inductor.