IP Library Granted Patent US 7,211,378
Granted Patent B2
US 7,211,378 · App. 10/353,856 · Granted May 1, 2007

Filovirus vectors and noninfectious filovirus-based particles

Assignee: Wisconsin Alumni Research Foundation
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Quick Facts
Patent No.
US 7,211,378
App. No.
10/353,856
Granted
May 1, 2007
Kind
B2
Abstract

Cloned filovirus genomic cDNA and methods of using the cDNA are provided. Further provided are noninfectious lipid encapsulated filovirus-based particles.

Claims (14)

1. A method to prepare lipid encapsulated replication-defective particles comprising filovirus matrix protein and a filovirus glycoprotein, comprising: a) providing a supernatant from a culture of eukaryotic cells contacted with a first DNA vector comprising a promoter operably linked to a DNA encoding a filovirus matrix protein or a portion thereof having residues corresponding to residues 1–226, 1–176, 50–326, or 100–326 of the Ebola virus matrix protein, which is capable of being incorporated into a particle, and a second DNA vector comprising a promoter operably linked to a DNA fragment of interest encoding a protein, wherein if the protein encoded by the DNA fragment of interest is not a glycoprotein, the culture is also contacted with a third DNA vector encoding a glycoprotein; and b) isolating from the supernatant replication-defective lipid encapsulated filamentous particles comprising filovirus matrix protein or the portion thereof, and the filovirus glycoprotein or the second DNA vector and the glycoprotein.

2. The method of claim 1 wherein the cell is a mammalian cell.

3. The method of claim 1 wherein the DNA fragment of interest encodes a therapeutic protein, an immunogenic protein or peptide of a pathogen, or a tumor antigen.

4. The method of claim 1 wherein the lipid encapsulated particles comprise a therapeutic protein, an immunogenic protein or peptide, or a tumor antigen, or a nucleic acid encoding the therapeutic protein, immunogenic protein or peptide, or tumor antigen.

5. The method of claim 1 wherein the DNA fragment encodes an integral membrane protein, a transmembrane protein, or a viral glycoprotein.

6. The method of claim 5 wherein the glycoprotein is a filovirus glycoprotein.

7. The method of claim 1 wherein the DNA fragment encodes a fusion protein.

8. The method of claim 5 wherein the viral glycoprotein is a chimeric protein.

9. The method of claim 1 wherein the matrix protein is Ebola virus VP40.

10. Lipid encapsulated particles prepared by the method of claim 1 .

11. The method of claim 1 wherein the DNA encoding the filovirus matrix protein comprises one or more nucleotide insertions, deletions or substitutions relative to DNA encoding wild-type filovirus matrix protein.

12. The method of claim 5 wherein the viral glycoprotein is a filovirus glycoprotein.

13. The method of claim 1 wherein codon 14 of the filovirus matrix protein encodes an alanine.

14. The method of claim 1 wherein the particles have a diameter of about 65 to 85 nm.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 18, 2003
From: YOSHIHIRO, KAWAOKA; NEUMANN, GABRIELE; JASENOSKY, LUKE D.
To: WISCONSIN ALUMNI RESEARCH FOUNDATION
Reel/Frame 013745/0226 →
Continuity (2)
Provisional Application 6035397200 · Jan 31, 2002
Related Publication 20030215794A1 · Nov 20, 2003