Apparatus and method for electroporation of biological samples
First Claim
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1. A flow electroporation device comprising:
- a chamber for containing a suspension of cells to be electroporated;
the chamber being at least partially defined by opposing oppositely chargeable electrodes; and
wherein the thermal resistance of the chamber is less than approximately 10°
C. per Watt.
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Abstract
The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations in blood by electroporation to achieve therapeutically desirable changes in the physical characteristics of the various cell populations in blood.
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Citations
116 Claims
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1. A flow electroporation device comprising:
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a chamber for containing a suspension of cells to be electroporated; the chamber being at least partially defined by opposing oppositely chargeable electrodes; and wherein the thermal resistance of the chamber is less than approximately 10°
C. per Watt. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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- 21. A method of electroporating a cell comprising flowing a suspension of cells to be electroporated through an electric field in a flow chamber, the electric field being produced by opposing oppositely charged electrodes at least partially defining the flow chamber, wherein the thermal resistance of the flow chamber is less than approximately 10°
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41. A flow electroporation device comprising:
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walls defining a flow channel having an electroporation zone configured to receive and to transiently contain a continuous flow of a suspension of cells to be electroporated; an inlet flow portal in fluid communication with the flow channel, whereby the suspension can be introduced into the flow channel through the inlet flow portal; an outlet flow portal in fluid conmiunication with the flow channel, whereby the suspension can be withdrawn from the flow channel through the outlet portal; the walls defining the flow channel within the electroporation zone comprising a first electrode forming a substantial portion of a first wall of the flow channel and a second electrode forming a substantial portion of a second wall of the flow channel opposite the first wall, the first and second electrodes being such that when placed in electrical communication with a source of electrical energy an electric field is formed therebetween through which the suspension can flow; and wherein the thermal resistance of the flow channel is less than approximately 10°
C. per Watt. - View Dependent Claims (42, 43, 44, 45, 46, 47)
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48. A flow electroporation device comprising:
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a flow chamber for containing a suspension of cells to be electroporated; the flow chamber being at least partially defined by opposing oppositely chargeable electrodes; and wherein the flow chamber has a ratio of combined electrode surface in contact with buffer to the distance between the electrodes of approximately 1 to 100 cm. - View Dependent Claims (49, 50, 51, 52, 53)
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54. A flow electroporation device comprising:
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walls defining a flow channel configured to receive and to transiently contain a continuous flow of a suspension of cells to be electroporated; an inlet flow portal in fluid communication with the flow channel, whereby the suspension can be introduced into the flow channel through the inlet flow portal; an outlet flow portal in fluid communication with the flow channel, whereby the suspension can be withdrawn from the flow channel through the outlet portal; the walls defining the flow channel comprising a first electrode forming at least a portion of a first wall of the flow channel and a second electrode forming at least a portion of a second wall of the flow channel opposite the first wall, the first and second electrodes being such that when placed in electrical communication with a source of electrical energy an electric field is formed therebetween through which the suspension can flow; and wherein the thermal resistance of the flow channel is less than approximately 10°
C. per Watt. - View Dependent Claims (55, 56, 57, 58, 59, 60, 61, 62, 63, 64)
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65. A flow electroporation device, comprising:
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walls defining a flow channel configured to receive and to transiently contain a continuous flow of a suspension comprising particles; an inlet flow portal in fluid communication with the flow channel, whereby the suspension can be introduced into the flow channel through the inlet flow portal; an outlet flow portal in fluid communication with the flow channel, whereby the suspension can be withdrawn from the flow channel through the outlet flow portal; the walls defining the flow channel comprising a first electrode plate forming a first wall of the flow channel and a second electrode plate forming a second wall of the flow channel opposite the first wall;
wherein the area of the electrodes contact with the suspension, and the distance between the electrodes is chosen so that the thermal resistance of the flow channel is less than approximately 4°
C. per Watt;the paired electrodes placed in electrical communication with a source of electrical energy, whereby an electrical field is formed between the electrodes; whereby the suspension of the particles flowing through the flow channel can be subjected to an electrical field formed between the electrodes. - View Dependent Claims (66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85)
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- 86. A method of transfecting a cell comprising providing a nucleic acid or an expression vector coding for a desired protein or peptide and introducing the expression vector into the cell by flow electroporation using a flow channel having a thermal resistance of less than approximately 10°
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109. A method of delivering a therapeutic agent to patient comprising:
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incorporating the therapeutic agent into platelets by electroporation using a flow electroporation channel having a thermal resistance of less than approximately 10°
C. per Watt; andadministering the platelets to the patient. - View Dependent Claims (110, 111, 112, 113, 114)
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115. A method comprising:
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flowing a suspension of CLL-B cells to be electroporated through a flow channel; and exposing said suspension of CLL-B cells to an electric field while flowing through said flow channel, said electric field having a strength of greater than 0.5 kV/cm. - View Dependent Claims (116)
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Specification