Cellectis S.a.

Cyto Pulse Sciences, Inc.

Other

Cellectis

Walters, Richard E.

King, Alan D.

Alan D King

Richard E Walters

Derin C Walters

Stephen B Deitz

Donald J Rodis, Jr

Anna-Karin Roos

Christoph Leder

Maxim Pavlenko

Pavel Pisa

Anna-Karin Maltais

An object of the invention is to provide an electroporation method for treating vesicles with exogenous material for insertion of the exogenous material into the vesicles which includes the steps of: a. retaining a suspension of the vesicles and the exogenous material in a treatment volume in a chamber which includes electrodes, wherein the chamber has a geometric factor (cm) defined by the quotient of the electrode gap squared (cm) divided by the chamber volume (cm), wherein the geometric factor is less than or equal to 0.1 cm, wherein the suspension of the vesicles and the exogenous material is in a medium which is adjusted such that the medium has conductivity in a range spanning 50 microSiemens/cm to 500 microSiemens/cm, wherein the suspension is enclosed in the chamber during treatment, and b. treating the suspension enclosed in the chamber with one or more pulsed electric fields. With the method, the treatment volume of the suspension is scalable, and the time of treatment of the vesicles in the chamber is substantially uniform.

Large volume ex vivo electroporation method

An electroporation apparatus and its novel chamber with inlet ports for mixing cells and exogenous material. The inlet ports are oriented in nonparallel to each other immediately adjacent at the same top corner of the first wall of the chamber. The mixing chamber comprises successive wall sections, two curved walls at its bottom; the first curved corner is on the same side of the chamber where the liquids enter the chamber, and directs the liquids to the second curved corner at the opposing side of the chamber which in turn further redirects the mixing to the first curved corner. The direction of the liquid flow mixture change direction at least twice into the mixing chamber.

Dynamic mixing and electroporation chamber and system

An object of the invention is to provide an electroporation method for treating vesicles with exogenous material for insertion of the exogenous material into the vesicles which includes the steps of: a. retaining a suspension of the vesicles and the exogenous material in a treatment volume in a chamber which includes electrodes, wherein the chamber has a geometric factor (cm.sup.&#x2212;1) defined by the quotient of the electrode gap squared (cm.sup.2) divided by the chamber volume (cm.sup.3), wherein the geometric factor is less than or equal to 0.1 cm.sup.&#x2212;1, wherein the suspension of the vesicles and the exogenous material is in a medium which is adjusted such that the medium has conductivity in a range spanning 50 microSiemens/cm to 500 microSiemens/cm, wherein the suspension is enclosed in the chamber during treatment, and b. treating the suspension enclosed in the chamber with one or more pulsed electric fields. With the method, the treatment volume of the suspension is scalable, and the time of treatment of the vesicles in the chamber is substantially uniform.

An object of the invention is to provide a method and apparatus for the delivery of polynucleotide vaccines into mammalian skin cells to increase T cell response and to reduce pain and discomfort due to long electric waveform application and due to muscle contractions. The method for the delivery of polynucleotide vaccines into mammalian skin cells includes the steps of: (a.) administering a polynucleotide vaccine into the skin at an administration site, (b.) applying a needle electrode to the skin in the vicinity to the administration site, and (c.) applying a sequence of at least three single, operator-controlled, independently programmed, narrow interval electrical waveforms, which have pulse intervals that are less than 100 milliseconds, to deliver the polynucleotide vaccine into the skin cells by electroporation. The sequence of at least three waveforms has one, two, or three of the following characteristics (1) at least two of the at least three waveforms differ from each other in waveform amplitude, (2) at least two of the at least three waveforms differ from each other in waveform width, and (3) a first waveform interval for a first set of two of the at least three waveforms is different from a second waveform interval for a second set of two of the at least three waveforms.

Method and apparatus for the delivery of polynucleotide vaccines to mammalian skin

An object of the invention is to provide methods and compositions relating to a vaccine against prostate cancer which includes a non-human-primate PSA for administration to humans to provide an immune response against human PSA. More generally, an object of the invention is to provide methods and compositions relating to using a non-human primate xenogeneic antigen (e.g. protein) in a human, wherein, with respect to the non-human primate xenogeneic antigen that is used, there are relatively few interspecies differences between the non-human primate xenogeneic antigen and the human self antigen in order to induce an optimal immune response in the human to its native self antigen.

Methods and compositions relating to a vaccine against prostate cancer

An object of the invention is to provide a method of treating biological cells prior to subjecting the biological cells to cell fusion pulses which includes the step of treating the biological cells with pre-fusion electric field waveforms which change amplitude in a non-linear way with respect to time, such that the biological cells are first aligned with a relatively low amplitude, long duration pre-fusion electric field waveform and then compressed with a relatively high amplitude, short duration pre-fusion electric field waveform resulting in increased cell membrane contact prior to being subjected to cell fusion. The non-linear pre-fusion electric field waveforms can change in a stepped way, in a continuous way, in a sigmoidal way, with step-wise increasing waveforms in adjacent steps, with step-wise increasing waveforms in non-adjacent steps, and in accordance with non-linear algorithms.

Non-linear amplitude dielectrophoresis waveform for cell fusion

An object of the invention is to provide a method for delivery of macromolecules into biological cells in the tissues of a patient and includes the steps of: (a) providing electrodes ( ) in an electrode assembly ( ), wherein the electrodes have fixed electrode surfaces ( ) which are coated with at least one static layer of electrode releasable molecules ( ) to be delivered; (b) providing a waveform generator ( ) for generating electric fields; (c) establishing electrically conductive pathways between the electrodes ( ) and the waveform generator ( ); (d) locating the electrodes ( ) such that the biological cells are situated therebetween, and (g) providing electric fields in the form of pulse waveforms from the waveform generator ( ) to the electrodes ( ), such that molecules in the at least one static layer of the electrode releasable molecules ( ) on the electrodes ( ) are delivered into the biological cells. The electrode releasable molecules ( ) can be either electric field separable molecules and/or solvent separable material. Another object of the invention is to provide an apparatus for carrying out the method of the invention. The static-coated electrode assembly ( ) can be provided in a sterile package ( ), from which the electrode assembly ( ) is removed prior to use. The statically-coated electrode assembly ( ) can be in a form of a disposable assembly ( ) which is removable and replaceable from an electrode assembly holder ( ).

Electrodes coated with treating agent and uses thereof

The object of the invention is to provide a method and apparatus for treating membrane-containing material with electrical fields, in vitro, and with an added treating substance. With the method, a plurality of electrodes are arrayed around the material to be treated and are connected to outputs of an electrode selection apparatus. Inputs of the electrode selection apparatus are connected to outputs of a pulse sequence generator. A treating substance is added to the membrane-containing material. Electrical pulses are applied to the electrode selection apparatus and are routed through the electrode selection apparatus in a predetermined, computer-controlled sequence to selected electrodes in the array of electrodes, whereby the membrane-containing material is treated with the added treating substance and with electrical fields. The routing of applied pulses through the electrode selection apparatus to selected electrodes can be done in an enormous number of ways.

Method and apparatus for treating materials with electrical fields having varying orientations

An object of the invention is to provide a method for delivery of macromolecules into biological cells, such as Langerhans cells ( ) in the epidermis ( ) of a patient, which includes the steps of coating electodes ( ) in an electrode assembly ( ) with solid phase macromolecules to be delivered, such as a DNA, and/or RNA vaccine or a protein-based vaccine, attaching the electrode assembly ( ) having the coated electrodes ( ) to an electrode assembly holder ( ), providing a waveform generator ( ), establishing electrically conductive pathways between the electrodes ( ), and the waveform generator ( ), locating the electrodes ( ) such that the biological cells are situated therebetween, such as by penetrating the needle electrode ( ) into the epidermis ( ) above the epidermal basal lamina, and providing pulse waveform from the waveform generator ( ) to the electrodes ( ), such that macromolecule on the electrodes ( ) is driven off of the electrodes ( ), and delivered into the biological cells, such as the Langerhans cells ( ).

Delivery of macromolecules into cells

Method and apparatus for treating materials with electrical fields

A method is provided for treating materials, especially organic materials, with pulsed electrical fields, wherein the method includes the step of applying an agile pulse sequence having at least three pulses to a material, wherein the agile pulse sequence has one, two, or three of the following characteristics: (1) at least two of the at least three pulses differ from each other in pulse amplitude; (2) at least two of the at least three pulses differ from each other in pulse width; and (3) a first pulse interval for a first set of two of the at least three pulses is different from a second pulse interval for a second set of two of the at least three pulses. When biological cells are treated to form pores in an electroporation procedure, the induced pores are sustained for a relatively long period of time, and viability of the biological cells is maintained.

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Highland, MD, United States of America

Alan D King