Transfection process

• US 5,766,902 A
• Filed: 06/07/1995
• Issued: 06/16/1998
• Est. Priority Date: 08/20/1993
• Status: Expired due to Fees

• Current Assignee: M.L. Laboratories, PLC (Vectura Group Ltd.)
• Original Assignee: The Therexsys Limited (Vectura Group Ltd.)
• Inventors: Djeha, Hakim, Antoniou, Mike, Craig, Roger Kingdon
• Primary Examiner(s): Ketter, James
• Assistant Examiner(s): Yucel, Irem
• Application Number:
  US08/487,473
• Time in Patent Office: 1,105 Days
• Field of Search: 514/44, 435/2, 435/240, 435/172.3, 435/173.5, 424/450, 935/52, 935/62, 530/391.7
• US Class Current: 435/461
• CPC Class Codes: A61K 47/6807 the drug or compound being a sugar, nucleoside, nucleotide, nucleic acid, e.g. RNA antisense
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

A method for delivering to a target cell in a population of cells a biologically active agent comprising the steps of exposing the population of cells to a complex comprising the biologically active agent and a ligand capable of binding to the target cell and subjecting the population of cells to an electric field while maintaining cell viability.

Claims

1. An in vitro method for delivering to a target cell in a population of cells a nucleic acid molecule, the method comprising the steps of:

a) exposing the population of cells to a complex comprising the nucleic acid molecule and a ligand which binds to the target cell; and

b) subjecting the population of cells to a mild electric field.

2. The method of claim 1 wherein the complex comprises an endosomal disruption agent.

3. The method of claim 2 wherein the endosomal disruption agent is an influenza virus haemagglutinin fusogenic peptide.

4. The method of claim 1 wherein the population of cells is heterogenous.

5. The method of claim 1 wherein the target cell is a cell of a subpopulation of said population of cells.

6. The method of claim 1 wherein the nucleic acid molecule encodes a protein or ribozyme.

7. The method of claim 1 wherein the nucleic acid molecule is a gene useful in therapy.

8. The method of claim 7 wherein the gene is useful in the correction of a genetic defect.

9. The method of claim 7 wherein the gene is useful in the therapy of an infectious disease.

10. The method of claim 9 wherein the infectious disease is AIDS.

11. The method of claim 10 wherein the nucleic acid molecule encodes a decoy protein or ribozyme.

12. The method of claim 7 wherein the gene is useful in the correction of a protein deficiency.

13. The method of claim 1 wherein the ligand is an antibody or a fragment thereof.

14. The method of claim 1 wherein the ligand is a growth factor.

15. The method of claim 1 wherein the target cell is a stem cell.

16. The method of claim 15 wherein the target cell is a haematopoietic stem cell.

17. The method of claim 1 wherein the target cell found in a population of hematopoietic stem cells.

18. The method of claim 17 wherein the target cell is a T cell.

19. An in vitro method of transporting across a cell membrane a biologically active agent, the method comprising

a) exposing the cell membrane of an intact cell in vitro to a complex comprising a nucleic acid and a ligand which binds to the target cell;

and

b) subjecting the cell membrane in vitro to a mild electric field.