Electrospraying apparatus and method for introducing material into cells

• US 6,093,557 A
• Filed: 06/05/1998
• Issued: 07/25/2000
• Est. Priority Date: 06/12/1997
• Status: Expired due to Term

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Chen, Da-Ren, Pui, David Y. H.
• Primary Examiner(s): Weber, Jon P.
• Application Number:
  US09/092,794
• Time in Patent Office: 781 Days
• Field of Search: 435/173.1, 435/285.3
• US Class Current: 435/173.1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: B01J 2/006 Coating of the granules without description of the process or the device by which the granules are obtained to prevent the granules sticking together B01J2/30
• CPC Class Codes: B01J 2/04 in a gaseous medium if combined with suspending the material in a gas, e.g. fluidised beds B01J2/16
• CPC Class Codes: B01J 2219/00371 comprising electrodes
• CPC Class Codes: B01J 2219/00436 Maskless processes
• CPC Class Codes: B05B 13/04 the spray heads being moved during spraying operation
• CPC Class Codes: B05B 5/025 Discharge apparatus, e.g. electrostatic spray guns
• CPC Class Codes: B05B 5/03 characterised by the use of gas , e.g. electrostatically assisted pneumatic spraying B05B5/04, B05B5/043, B05B5/047 take precedence
• CPC Class Codes: B05B 5/0533 Electrodes specially adapted therefor; Arrangements of electrodes
• CPC Class Codes: B05B 5/081 specially adapted for treating particulate materials
• CPC Class Codes: B05B 5/082 characterised by means for supporting, holding or conveying the objects
• CPC Class Codes: B05B 5/084 the objects lying on, or being supported above conveying means, e.g. conveyor belts
• CPC Class Codes: B05B 5/087 Arrangements of electrodes, e.g. of charging, shielding, collecting electrodes B05B5/12, B05B5/14 take precedence; arrangements of electrodes on the discharge apparatus B05B5/0533
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 15/895 using biolistic methods

Abstract

A method of introducing biological material into cells includes providing one or more target cells and establishing a spray of substantially dispersed particles including biological material. The substantially dispersed particles have an electrical charge applied thereto such that one or more of the substantially dispersed particles of the spray is introduced into one or more of the target cells. The spray of substantially dispersed particles may be established by dispensing a spray of microdroplets suspending particles. The electrical charge is concentrated on the suspended particles as the microdroplet evaporates. The suspended particles may include carrier particles with biological material or the suspended particles may be particles of biological material alone. The space charge effect of the concentrated electrical charge on the substantially dispersed particles of the spray enable one or more of the particles to be introduced into one or more of the target cells.

Claims

1. A method of introducing biological material into cells, the method comprising:

providing one or more target cells; and

establishing a spray of substantially dispersed particles including at least biological material, the substantially dispersed particles having an electrical charge applied thereto such that a space charge effect resulting from repulsion of the electrically charged substantially dispersed particles is used to provide one or more of the substantially dispersed particles of the spray with a velocity sufficient for introduction of the one or more substantially dispersed particles into one or more of the target cells.

2. The method according to claim 1, wherein the step of establishing the spray of substantially dispersed particles includes dispensing a spray of microdroplets suspending one or more particles, and further wherein the electrical charge is concentrated on the particles suspended by the microdroplets as the microdroplets evaporates.

3. The method according to claim 2, wherein the suspended particles include carrier particles and biological material.

4. The method according to claim 3, wherein the microdroplets have a nominal diameter of about 10 nanometers to about 10 microns and the carrier particles have a nominal diameter of about 2 nanometers to about 1 micron.

5. The method according to claim 2, wherein the suspended particles are particles of biological material.

6. The method according to claim 2, wherein the step of dispensing the spray of microdroplets suspending particles includes creating a nonuniform electrical field between a dispensing tip from which the spray is established and an electrode electrically isolated from the dispensing tip.

7. The method according to claim 6, wherein the method further includes the step of directing the spray of substantially dispersed particles towards the one or more target cells using the electrode isolated from the dispensing tip.

8. The method according to claim 2, wherein the space charge effect of the concentrated electrical charge on the substantially dispersed particles of the spray enable one or more of the particles to penetrate one or more of the target cells.

9. The method according to claim 8, wherein the electrical charge concentrated on a particular particle suspended by a certain microdroplet is in the range of about 80 percent to about 95 percent of a maximum charge that can be held by the certain microdroplet suspending the particular particle.

10. The method according to claim 1, wherein the step of establishing a spray of substantially dispersed particles includes establishing a continuous spray of substantially dispersed particles.

11. The method according to claim 1, wherein the step of establishing a spray of substantially dispersed particles includes dispensing a spray of powdered biological material.

12. A method for introducing biological material into cells, the method comprising:

providing one or more target cells;

providing a first flow of a suspension including at least biological material;

providing a second flow of a solution; and

establishing a spray of substantially dispersed particles including at least biological material from the first flow and the second flow, the substantially dispersed particles having an electrical charge applied thereto such that a space charge effect resulting from repulsion of the electrically charged substantially dispersed particles is used to provide one or more of the substantially dispersed particles of the spray with a velocity sufficient for introduction of the one or more substantially dispersed particles into one or more of the target cells.

13. The method according to claim 12, wherein the step of establishing the spray of substantially dispersed particles includes dispensing a spray of microdroplets suspending one or more particles, and further wherein the electrical charge is concentrated on the particles suspended by the microdroplets as the microdroplets evaporates.

14. The method according to claim 13, wherein the suspended particles include carrier particles and biological material.

15. The method according to claim 13, wherein the suspended particles consist essentially of biological material.

16. The method according to claim 12, wherein the solution includes an agent for promoting penetration of the target cells.

17. The method according to claim 12, wherein the solution has a predetermined electrical conductivity, wherein the predetermined electrical conductivity of the solution is selected to control the space charge effect.