公开(公告)号：US06916656B2

公开(公告)日：2005-07-12

申请号：US10487235

申请日：2002-04-05

申请人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King

发明人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King

IPC分类号： C12N15/02 ,  C12N5/10 ,  C12N5/16 ,  C12N13/00 ,  C12N15/64 ,  C12N15/74 ,  C12N15/87

CPC分类号： C12M35/02 ,  A61K2039/5152 ,  A61K2039/5154 ,  C12N5/16 ,  C12N13/00 ,  H01L2224/83856 ,  H01L2924/01019

摘要： 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.

摘要翻译： 本发明的一个目的是提供一种在使生物细胞经历细胞融合脉冲之前处理生物细胞的方法，其包括用以非线性方式改变幅度的融合前电场波形来处理生物细胞的步骤， 相对于时间，使得生物细胞首先用相对低幅度，长持续时间的预融合电场波形对准，然后用相对较高幅度，短持续时间的预融合电场波形压缩，导致增加的细胞膜接触先 进行细胞融合。 非线性预融合电场波形可以以连续的方式以S形方式以相邻步骤中逐步增加的波形以逐步增加的波形在非相邻步骤中逐步增加，并以 按照非线性算法。

公开(公告)号：US20110282265A1

公开(公告)日：2011-11-17

申请号：US12451536

申请日：2008-05-20

申请人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King ,  Anna-Karin Roos

发明人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King ,  Anna-Karin Roos

IPC分类号： A61M37/00

CPC分类号： A61N1/327 ,  A61K39/12 ,  A61K2039/53 ,  A61K2039/54 ,  A61K2039/70 ,  A61N1/306 ,  C07K14/005 ,  C12N2770/24122 ,  C12N2770/24134

摘要： 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 reduce pain and discomfort due to long electric waveform application and due to muscle contractions. The method includes 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 15 polynucleotide vaccine into the skin cells by electroporation.

摘要翻译： 本发明的目的是提供一种用于将多核苷酸疫苗递送到哺乳动物皮肤细胞中以增加T细胞应答并减少由于长电波形应用引起的疼痛和不适以及由于肌肉收缩而引起的不适的方法和装置。 该方法包括在给药部位向皮肤施用多核苷酸疫苗，（b）将针电极施用于给药部位附近的皮肤，以及（c）施用至少三个单独的，操作者控制的， 具有脉冲间隔小于100毫秒的独立编程的窄间隔电波形，以通过电穿孔将15个多核苷酸疫苗递送到皮肤细胞中。

公开(公告)号：US6010613A

公开(公告)日：2000-01-04

申请号：US569664

申请日：1995-12-08

申请人： Richard E. Walters ,  Alan D. King

发明人： Richard E. Walters ,  Alan D. King

IPC分类号： C12N13/00

CPC分类号： C12N13/00 ,  C12M35/02

摘要： 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.

公开(公告)号：US06713291B2

公开(公告)日：2004-03-30

申请号：US09920861

申请日：2001-08-03

申请人： Alan D. King ,  Richard E. Walters

发明人： Alan D. King ,  Richard E. Walters

IPC分类号： C12N1300

CPC分类号： A61N1/325 ,  A61H39/002 ,  A61N1/306

摘要： 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 (16) in an electrode assembly (12), wherein the electrodes have fixed electrode surfaces (42) which are coated with at least one static layer of electrode releasable molecules (44) to be delivered; (b) providing a waveform generator (15) for generating electric fields; (c) establishing electrically conductive pathways between the electrodes (16) and the waveform generator (15); (d) locating the electrodes (16) such that the biological cells are situated therebetween, and (g) providing electric fields in the form of pulse waveforms from the waveform generator (15) to the electrodes (16), such that molecules in the at least one static layer of the electrode releasable molecules (44) on the electrodes (16) are delivered into the biological cells. The electrode releasable molecules (44) 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 (12) can be provided in a sterile package (24), from which the electrode assembly (12) is removed prior to use. The statically-coated electrode assembly (12) can be in a form of a disposable assembly (12) which is removable and replaceable from an electrode assembly holder (13).

公开(公告)号：US06653114B2

公开(公告)日：2003-11-25

申请号：US10265133

申请日：2002-10-07

申请人： Richard E. Walters ,  Alan D. King ,  Derin C. Walters

发明人： Richard E. Walters ,  Alan D. King ,  Derin C. Walters

IPC分类号： C12N1300

CPC分类号： C12N13/00 ,  C12M35/02

摘要： 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.

公开(公告)号：US09982251B2

公开(公告)日：2018-05-29

申请号：US10537254

申请日：2004-03-15

申请人： Richard E. Walters ,  Alan D. King

发明人： Richard E. Walters ,  Alan D. King

IPC分类号： C12N13/00 ,  C12N15/87 ,  C12M3/02 ,  C12M3/00 ,  C12M1/42 ,  C12M1/00 ,  A61K9/00

CPC分类号： C12N13/00 ,  A61K9/0009 ,  C12M1/42 ,  C12M3/00 ,  C12M3/006 ,  C12M3/02 ,  C12M35/02 ,  C12M45/07 ,  C12N15/87

摘要： 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.−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.−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.

公开(公告)号：US06603998B1

公开(公告)日：2003-08-05

申请号：US09889581

申请日：2001-07-30

申请人： Alan D. King ,  Richard E. Walters

发明人： Alan D. King ,  Richard E. Walters

IPC分类号： A61N130

CPC分类号： A61N1/325 ,  A61N1/306 ,  C12M35/02 ,  C12N15/87

摘要： An object of the invention is to provide a method for delivery of macromolecules into biological cells, such as Langerhans cells (22) in the epidermis (20) of a patient, which includes the steps of coating electodes (16) in an electrode assembly (12) with solid phase macromolecules to be delivered, such as a DNA, and/or RNA vaccine or a protein-based vaccine, attaching the electrode assembly (12) having the coated electrodes (16) to an electrode assembly holder (13), providing a waveform generator (15), establishing electrically conductive pathways between the electrodes (16), and the waveform generator (15), locating the electrodes (16) such that the biological cells are situated therebetween, such as by penetrating the needle electrode (16) into the epidermis (20) above the epidermal basal lamina, and providing pulse waveform from the waveform generator (15) to the electrodes (16), such that macromolecule on the electrodes (16) is driven off of the electrodes (16), and delivered into the biological cells, such as the Langerhans cells (22).

摘要翻译： 本发明的目的是提供一种用于将大分子递送到生物细胞的方法，例如患者的表皮（20）中的朗格汉斯细胞（22），其包括在电极组件中涂覆电极（16）的步骤 将具有涂覆电极（16）的电极组件（12）附接到电极组件保持器（13）上的要传递的固相大分子如DNA和/或RNA疫苗或基于蛋白质的疫苗， 提供波形发生器（15），在电极（16）和波形发生器（15）之间建立导电通路，定位电极（16），使得生物细胞位于其间，例如穿过针电极（ 16）进入表皮基底层上方的表皮（20），并将波形发生器（15）的脉搏波形提供给电极（16），使得电极（16）上的大分子从电极（16）驱动， ，并交付 生物细胞，如朗格汉斯细胞（22）。

公开(公告)号：US6117660A

公开(公告)日：2000-09-12

申请号：US242174

申请日：1999-02-10

申请人： Richard E. Walters ,  Alan D. King ,  Derin C. Walters

发明人： Richard E. Walters ,  Alan D. King ,  Derin C. Walters

IPC分类号： C12N13/00

CPC分类号： C12N13/00 ,  C12M35/02

摘要： The object of the invention is to provide a method and apparatus for treating membrane containing material with electrical fields and with an added treating substance. With the method, a plurality of electrodes (121-128) are arrayed around the material to be treated and are connected to outputs of an electrode selection apparatus (110). Inputs of the electrode selection apparatus are connected to outputs of an agile 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 of sequentially varying directions. The routing of applied pulses through the electrode selection apparatus (110) to selected electrodes (121-128) can be done in an enormous number of ways.

摘要翻译： PCT No.PCT / US97 / 09300 Sec。 371日期1999年2月10日 102（e）1999年2月10日PCT 1997年6月10日PCT公布。 第WO98 / 56893号公报 日期：1998年12月17日本发明的目的是提供一种用电场和附加处理物质处理含膜材料的方法和装置。 利用该方法，将多个电极（121-128）围绕待处理的材料排列并连接到电极选择装置（110）的输出端。 电极选择装置的输入连接到敏捷脉冲序列发生器的输出端。 将处理物质加入到含膜材料中。 将电脉冲施加到电极选择装置，并以预定的计算机控制的顺序通过电极选择装置引导到电极阵列中的选定电极，由此将含有膜的材料用添加的处理物质和电场 顺序变化的方向。 施加的脉冲通过电极选择装置（110）到选定电极（121-128）的布线可以以许多方式进行。

公开(公告)号：US20110166501A1

公开(公告)日：2011-07-07

申请号：US13065135

申请日：2011-03-15

申请人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King ,  Anna-Karin Roos

发明人： Richard E. Walters ,  Derin C. Walters ,  Alan D. King ,  Anna-Karin Roos

IPC分类号： A61M37/00

CPC分类号： A61N1/327 ,  A61K39/12 ,  A61K2039/53 ,  A61K2039/54 ,  A61K2039/70 ,  A61N1/306 ,  C07K14/005 ,  C12N2770/24122 ,  C12N2770/24134

摘要： 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.

摘要翻译： 本发明的目的是提供一种用于将多核苷酸疫苗递送到哺乳动物皮肤细胞中以增加T细胞应答并减少由于长电波形应用引起的疼痛和不适以及由于肌肉收缩引起的疼痛和不适的方法和装置。 用于将多核苷酸疫苗递送到哺乳动物皮肤细胞中的方法包括以下步骤：（a）在给药部位将多核苷酸疫苗施用于皮肤，（b）将针电极施用于给药的附近的皮肤 站点，以及（c）应用脉冲间隔小于100毫秒的至少三个单个，操作者控制的，独立编程的窄间隔电波形序列，以通过电穿孔将多核苷酸疫苗递送到皮肤细胞中 。 至少三个波形的序列具有以下特征（1）中的一个，两个或三个，所述至少三个波形中的至少两个波形在波形幅度上彼此不同，（2）至少三个波形中的至少两个波形 在波形宽度上彼此不同，以及（3）至少三个波形中的两个的第一组的第一波形间隔与用于至少三个波形中的两个的第二组的第二波形间隔不同。

公开(公告)号：US20110000550A1

公开(公告)日：2011-01-06

申请号：US12761351

申请日：2010-04-15

申请人： Stephen J. Rogier ,  Lyman W. Fawcett, JR. ,  Alan D. King

发明人： Stephen J. Rogier ,  Lyman W. Fawcett, JR. ,  Alan D. King

IPC分类号： F16K15/20

CPC分类号： B63C9/18 ,  B63C9/19 ,  Y10T137/0318 ,  Y10T137/1684 ,  Y10T137/3662 ,  Y10T137/6123 ,  Y10T137/8359

摘要： A manual inflator having a status indicator that indicates whether a fully-charged, unspent gas cylinder has been installed on the inflator. The status indicator comprises a “single point” indicator having an indicator window that displays the color “green” when the automatic inflator is fully operational or the color “red” when the inflator is at least partially inoperable automatically due to the removal of the gas cylinder or due to the firing of the inflator resulting in a spent gas cylinder.

摘要翻译： 具有状态指示器的手动充气机，其指示充气机上是否安装了充满电的未充气气瓶。 状态指示器包括“单点”指示器，当自动充气机完全运行时，具有显示颜色“绿色”的指示器窗口，或者当充气机由于气体的去除而自动至少部分地不能操作时，颜色为“红色” 或者由于充气机的点火导致废气瓶。