Minicell libraries

• US 20030198996A1
• Filed: 05/28/2002
• Published: 10/23/2003
• Est. Priority Date: 05/24/2001
• Status: Abandoned Application

• Current Assignee: Mpex Biosciences, Inc.
• Original Assignee: Mpex Biosciences, Inc.
• Inventors: Berkley, Neil, Sabbadini, Roger A., Surber, Mark W., Gerhart, William
• Application Number:
  US10/157,147
• Publication Number:
  US 20030198996A1
• Time in Patent Office: Days
• US Class Current: 506/14
• 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 library, comprising two or more minicells, wherein each minicell comprises a different exogenous protein.

2. The minicell library of claim 1, wherein said minicell is a eubacterial minicell, a poroplast, a spheroplast or a protoplast.

3. The minicell library of claim 1, wherein said exogenous protein is a displayed protein.

4. The minicell library of claim 1, wherein said exogenous protein is a membrane protein.

5. The minicell library of claim 4, wherein said membrane protein is a receptor.

6. The minicell library of claim 1, wherein said protein is a soluble protein that is contained within or secreted from said minicell.

7. The minicell library of claim 1, wherein minicells within said library comprise an expression element that comprises expression sequences operably linked to a nucleic acid having an ORF that encodes said exogenous protein.

8. The minicell library of claim 7, wherein said nucleic acid has been mutagenized.

9. The minicell library of claim 7, wherein an active site of said exogenous protein has a known or predicted three-dimensional structure, and said a portion of said ORF encoding said active site has been mutagenized.

10. The minicell library of claim 1, wherein each of said minicells comprises an exogenous protein that is a variant of a protein having a known or predicted three-dimensional structure.

11. A minicell library, comprising two or more minicells, wherein each minicell comprises a different fusion protein, each of said fusion protein comprising a first polypeptide that is a constant polypeptide, wherein said constant polypeptide comprises at least one transmembrane domain or at least one membrane anchoring domain, and a second polypeptide, wherein said second polypeptide is a variable amino acid sequence that is different in each fusion proteins.

12. The minicell library of claim 11, wherein minicells within said library comprise an expression element that comprises expression sequences operably linked to a nucleic acid having an ORF that encodes said fusion protein.

13. The minicell library of claim 12, wherein said second polypeptide of said fusion protein is encoded by a nucleic acid that has been cloned.

14. The minicell library of claim 12, wherein each of said second polypeptide of each of said fusion proteins comprises a variant of an amino acid sequence from a protein having a known or predicted three-dimensional structure.

15. A minicell library, comprising-two or more minicells, wherein each minicell comprises a constant protein that is present in each minicell and a variable protein that differs from minicell to minicell.

16. The minicell library of claim 15, wherein one of said constant and variable proteins is a receptor, and the other of said constant and variable proteins is a co-receptor.

17. The minicell library of claim 15, wherein each of said constant and variable proteins is different from each other and is a factor in a signal transduction pathway.

18. The minicell library of claim 15, wherein one of said constant and variable proteins is a G-protein, and the other of said constant and variable proteins is a G-protein coupled receptor.

19. The minicell library of claim 15, wherein one of said constant and variable proteins comprises a first transrepression domain, and the other of said constant and variable comprises a second transrepression domain, wherein said transrepression domains limit or block expression of a reporter gene when said constant and variable proteins associate with each other.

20. The minicell library of claim 15, wherein one of said constant and variable proteins comprises a first transactivation domain, and the other of said constant and variable comprises a second transactivation domain, wherein said transactivation domains stimulate expression of a reporter gene when said constant and variable proteins associate with each other.