Richard W Walters, 68Fort Collins, CO

6603998, Aug 5, 2003

Jul 30, 2001

09/889581

Alan D. King - Takoma Park MD
Richard E. Walters - Columbia MD

Cyto Pulse Sciences, Inc. - Hanover MD

A61N 130

604 20, 604501

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 ( ).

6653114, Nov 25, 2003

Oct 7, 2002

10/265133

Richard E. Walters - Columbia MD, 21045
Alan D. King - Takoma Park MD, 20912
Derin C. Walters - Austin TX, 78758-3321

C12N 1300

4351736, 4351734, 4351735, 205701

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.

6713291, Mar 30, 2004

Aug 3, 2001

09/920861

Alan D. King - Takoma Park MD, 20912
Richard E. Walters - Columbia MD, 21045

C12N 1300

4351736

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.

6878538, Apr 12, 2005

Oct 28, 2003

10/694344

Richard E. Walters - Columbia MD, US
Benjamin R. Lane - Phoenix MD, US
Michael S. Abbott - Baltimore MD, US

Cyto Pulse Sciences, Inc. - Glen Burnie MD

C12M001/42

4352852, 4352873, 4352884, 4353052, 4353053, 435809, 20440301, 204412, 204400

An apparatus is provided for receiving and aligning a multiple electrode pair array and a multiple well plate used in electroporation. The apparatus includes a housing which includes a bottom housing portion and a pair of side housing portions which project upward from the bottom housing portion. A pair of board-reception structures are adjacent to the inner portions of the side housing portions. Multiple well plate engaging and disengaging means, supported by the housing, are provided for engaging a multiple well plate with a multiple electrode pair array and for disengaging a multiple well plate from a multiple electrode pair array. Preferably, the multiple well plate engaging and disengaging means include a multiple well plate lifting and lowering assembly. The multiple well plate lifting and lowering assembly can include lifting/lowering handles located outside the housing.

6916656, Jul 12, 2005

Apr 5, 2002

10/487235

Richard E. Walters - Columbia MD, US
Derin C. Walters - Columbia MD, US
Alan D. King - Highland MD, US

Cyto Pulse Sciences, Inc. - Glen Burnie MD

C12N015/02
C12N015/64

435450, 435461, 435470, 435471

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.

7371561, May 13, 2008

Oct 28, 2003

10/694345

Richard E. Walters - Columbia MD, US
Paul K. Gustavson - Gambrills MD, US

Cyto Pulse Sciences - Glen Burnie MD

C12M 1/42

4352852, 4352884, 4353052, 422 8202, 422102, 324450, 324692

A multiple electrode pair array apparatus is provided for use with a multiple well plate which has multiple wells distributed in a two-dimensional matrix array which has R rows and C columns. The multiple electrode pair array apparatus includes a non-conductive base member. An array of pairs of electrodes are attached to the base member and project therefrom. The pairs of electrodes are distributed on the base member in a two-dimensional matrix array which has R rows and C columns to enable registration with the two-dimensional matrix array of wells. Each pair of electrodes includes a respective first electrode and a respective second electrode. An array of R row conductors are electrically connected to corresponding first electrodes. An array of C column conductors are oriented perpendicular to the R row conductors, are electrically insulated from the row conductors, and are electrically connected to corresponding second electrodes.

7824901, Nov 2, 2010

Dec 1, 2003

10/580429

Richard E. Walters - Columbia MD, US

Cyto Pulse Sciences, Inc. - Glen Burnie MD

C12M 3/00
C12M 1/34

4352885

An apparatus is provided for carrying out fusion of biological cells () and includes a base member () on which a conductive outer electrode () is supported and has an outer electrode radius (r) and has an electrode height (). A conductive inner electrode () is supported on the base member () and has an inner electrode radius (r) and also has the electrode height (). The outer and inner electrodes () are spaced apart from each other by a gap which defines a fusion chamber (). The inner electrode radius (r), the outer electrode radius (r), and the gap are selected in accordance with a predetermined range of selectable ratios (r/r) in a range from 0. 7 to 0. 9, wherein a selected gap is limited by the range of selectable ratios (r/r), and wherein a determined ratio (r/r) among the selectable ratios is based on the selected gap, such that compression between the biological cells () and permeability between cell membranes are maximized and temperature rise is minimized for providing cell fusion in the fusion chamber ().

2004020, Oct 14, 2004

Mar 29, 2004

10/810829

Alan King - Takoma Park MD, US
Richard Walters - Columbia MD, US

C12N013/00
C12N015/01

435/173600, 435/446000

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 ().