Flow electroporation chamber and method

• US 20050019311A1
• Filed: 05/14/2004
• Published: 01/27/2005
• Est. Priority Date: 03/10/1995
• Status: Abandoned Application

• Current Assignee: Doug Doerfler, John W. Holaday, Peter H. Meserol, Shawn J. Green, Vininder Singh
• Original Assignee: Doug Doerfler, John W. Holaday, Peter H. Meserol, Shawn J. Green, Vininder Singh
• Inventors: Singh, Vininder, Meserol, Peter H., Doerfler, Doug, Holaday, John W., Green, Shawn J.
• Application Number:
  US10/846,729
• Publication Number:
  US 20050019311A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61K 35/18 Erythrocytes haemoglobin A61K38/42
• CPC Class Codes: A61M 1/16 with membranes
• CPC Class Codes: A61M 1/3687 Chemical treatment A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2202/0092 starting product created by centrifuging
• CPC Class Codes: A61M 2202/0429 Red blood cells; Erythrocytes
• CPC Class Codes: A61M 2205/3306 Optical measuring means
• CPC Class Codes: A61M 2205/3368 Temperature
• CPC Class Codes: A61M 2205/3382 Upper level detectors
• CPC Class Codes: A61M 2205/3606 cooled
• CPC Class Codes: A61M 2205/3673 thermo-electric, e.g. Peltier effect, thermocouples, semi-conductors
• CPC Class Codes: A61M 2205/50 with microprocessors or computers
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: F25B 21/02 using Peltier effect; using Nernst-Ettinghausen effect

Abstract

The present invention relates to a method and apparatus for the encapsulation of substances and drugs into cells and platelets. The present invention is also related to the incorporation of thrombus dissolving drugs, such as tissue plasminogen activator and streptokinase into platelets using the apparatus described herein. The treated platelets can then be used to treat patients suffering from a thrombus blocking a blood vessel. The present invention is also related to a preparation of red blood cells that has a stable right shift of the oxygen dissociation curve.

Claims

1-13. -13. Canceled

14. A method of incorporating a biologically-active substance into cells in a continuous flow system, comprising:

a. introducing the cells into the continuous flow system;

b. isolating the cells from at least some matter with which the cells are associated;

c. mixing the cells with a biologically-active substance to provide a cell suspension;

d. moving the cell suspension within the continuous flow system at a flow rate established by a computer-controllable pump to correspond to a sample processing rate;

e. electroporating the cell suspension, thereby causing the biologically-active substance to be encapsulated in the cells; and

f. incubating the cells to allow the cells to reseal to provide modified cells.

15. The method of claim 14, further comprising:

g. washing the modified cells to remove unencapsulated biologically-active substance therefrom.

16. The method of claim 14, the sample processing rate corresponding to a processing rate of a centrifuge.

17. The method of claim 14, the sample processing rate corresponding to an interval between pulses of electrical energy to the electrodes.

19. The method of claim 17, further comprising:

g. washing the modified cells to remove unencapsulated biologically-active substance therefrom.

21. The method of claim 19, the method comprising rotating a spindle to stretch an electrode wrapped around the spindle so as to break the electrode.

18. A method of incorporating a biologically-active substance into cells in a continuous flow system, comprising:

a. introducing the cells into the continuous flow system;

b. isolating the cells from at least some matter with which the cells are associated;

c. mixing the cells with a biologically-active substance to provide a cell suspension;

d. electroporating the cell suspension, thereby causing the biologically-active substance to be encapsulated in the cells;

e. controlling individual thermoelectric cooling elements to cool portions of the continuous flow system; and

f. incubating the cells to allow the cells to reseal to provide modified cells.

20. A method for rendering an electroporation apparatus non-operational, the method comprising automatically breaking an electrode prior to re-use.