Minicell-based delivery agents

• US 20030211599A1
• Filed: 05/28/2002
• Published: 11/13/2003
• Est. Priority Date: 05/24/2001
• Status: Abandoned Application

• Current Assignee: Mpex Biosciences, Inc.
• Original Assignee: Mpex Biosciences, Inc.
• Inventors: Klepper, Robert, Sabbadini, Roger A., Surber, Mark W.
• Application Number:
  US10/157,106
• Publication Number:
  US 20030211599A1
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/74 Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: C40B 40/02 Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/5432 Liposomes or microcapsules
• CPC Class Codes: G01N 33/60 involving radioactive labelled substances
• CPC Class Codes: Y02A 90/10 Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

The invention provides compositions and methods for the production of achromosomal and anucleate cells useful for applications such as diagnositic and therapeutic uses, as well as research tools and agents for drug discovery.

Claims

1. A minicell comprising a biologically active compound, wherein said minicell displays a ligand or binding moiety, wherein said ligand or binding moiety is part of a fusion protein comprising a first polypeptide that comprises at least one transmembrane domain or at least one membrane anchoring domain and a second polypeptide that comprises a binding moiety, and said minicell is a poroplast, spheroplast or protoplast.

3. The minicell of claim 1, wherein said binding moiety is selected from the group consisting of an antibody, an antibody derivative, a receptor and an active site of a non-catalytic derivative of an enzyme.

4. The minicell of claim 1, wherein said binding moiety is a single-chain antibody.

5. The minicell of claim 1, wherein said binding moiety is directed to a ligand selected from the group consisting of an epitope displayed on a pathogen, an epitope displayed on an infected cell and an epitope displayed on a hyperproliferative cell.

6. The minicell of claim 1, wherein said ligand is directed to a receptor selected from the group consisting of an epitope displayed on a pathogen, and epitope displayed on an infected cell and an epitope displayed on a hyperproliferative cell.

7. The minicell of claim 1, wherein said biologically active compound is selected from the group consisting of a radioisotope, a polypeptide, a nucleic acid and a small molecule.

8. The minicell of claim 1, further comprising a first and second nucleic acid, wherein said first nucleic acid comprises eukaryotic expression sequences operably linked to a first ORF, and a second nucleic acid, wherein said second nucleic acid comprises eubacterial expression sequences operably linked to a second ORF.

9. The minicell of claim 8, wherein one of said ORFs encodes a protein that comprises said binding moiety.

10. The minicell of claim 8, wherein said eubacterial expression sequences are induced and/or derepressed when said binding moiety is in contact with a target cell.

11. The minicell of claim 8, wherein said eukaryotic expression sequences are induced and/or derepressed when said nucleic acid is in the cytoplasm of a eukaryotic cell.

12. The minicell of claim 8, wherein the protein encoded by said first ORF comprises eukaryotic secretion sequences and/or the protein encoded by said second ORF comprises eubacterial secretion sequences.

2. A eubacterial minicell comprising a biologically active compound, wherein said minicell displays a binding moiety, wherein said binding moiety is selected from the group consisting of (a) a eukaryotic membrane protein;

(b) an archeabacterial membrane protein;

(c) an organellar membrane protein; and

(d) a fusion protein, said fusion protein comprising a first polypeptide, said first polypeptide comprising at least one transmembrane domain or at least one membrane anchoring domain; and

a second polypeptide, wherein said second polypeptide is not derived from a eubacterial protein and is neither a His tag nor a glutathione-S-transferase polypeptide, and wherein said polypeptide comprises a binding moiety.