# US 2004/0101929 A1 · Expressing mammalian protein complexes in fish

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- Number: US 2004/0101929 A1
- Document ID: US20040101929A1
- Status: Published application
- Kind: A1
- Application number: 10/302,570
- Filed: 2002-11-22
- Published: 2004-05-27
- Earliest claimed: 2002-11-22
- Inventors: Chi-Yao Chang; Yu-Shen Lai; Joseph Abraham Christopher John
- Web page: https://patentgrep.com/patent/US20040101929A1

## CPC

- C12N 15/8509: {for producing genetically modified animals, e.g. transgenic}
- A01K 67/0275: Genetically modified vertebrates, e.g. transgenic
- C07K 14/005: from viruses
- C07K 16/10: RNA viruses
- C12N 5/0618: {Cells of the nervous system}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A01K 2217/05: Animals comprising random inserted nucleic acids (transgenic)
- A01K 2227/40: Fish
- A01K 2267/01: Animal expressing industrially exogenous proteins
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- C12N 2770/30022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/30051: Methods of production or purification of viral material
- C12N 2840/20: translation of more than one cistron
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

A fish cell expressing a mammalian protein complex having two peptide species. The cell has a first nucleic acid sequence encoding a first peptide species of the mammalian protein complex and a second nucleic acid sequence encoding a second peptide species of the mammalian protein complex. A multiplicity of peptides of the first peptide species and the second peptide species are expressed and at least one peptide of the first peptide species and at least one peptide of the second peptide species assembly to form a functional mammalian protein complex. The invention also discloses a method of making such a cell, and a cultured fish cell infected with a virus or its progeny for producing the virus.

## Claims

### Claim 1

A fish cell that expresses a mammalian protein complex having two peptide species, the cell comprising:
a first nucleic acid sequence encoding a first peptide species of the mammalian protein complex, and
a second nucleic acid sequence encoding a second peptide species of the mammalian protein complex, wherein a multiplicity of peptides of the first peptide species and the second peptide species are expressed and at least one peptide of the first peptide species and at least one peptide of the second peptide species assembly to form a functional mammalian protein complex.

### Claim 2 (depends on 1)

The cell of claim 1, wherein the mammalian protein complex is an antibody.

### Claim 3 (depends on 2)

The cell of claim 2 is a brain cell.

### Claim 4 (depends on 2)

The cell of claim 2, wherein the antibody contains a disulfide bond.

### Claim 5 (depends on 2)

The cell of claim 2, wherein the fish is a yellow grouper.

### Claim 6 (depends on 5)

The cell of claim 5 is a brain cell.

### Claim 7 (depends on 2)

The cell of claim 2, wherein the antibody binds to a microorganism.

### Claim 8 (depends on 7)

The cell of claim 7, wherein the fish is a yellow grouper.

### Claim 9 (depends on 7)

The cell of claim 7, wherein the microorganism is a virus.

### Claim 10 (depends on 9)

The cell of claim 9, wherein the fish is a yellow grouper.

### Claim 11 (depends on 9)

The cell of claim 9, wherein the virus is a nervous necrosis virus.

### Claim 12 (depends on 11)

The cell of claim 11, wherein the fish is a yellow grouper.

### Claim 13 (depends on 12)

The cell of claim 12, wherein the antibody binds to a coat protein of the virus.

### Claim 14 (depends on 13)

The cell of claim 13 is a brain cell.

### Claim 15 (depends on 1)

The cell of claim 1, wherein the mammalian protein complex contains a disulfide bond.

### Claim 16 (depends on 15)

The cell of claim 15 is a brain cell.

### Claim 17 (depends on 15)

The cell of claim 15, wherein the fish is a yellow grouper.

### Claim 18 (depends on 17)

The cell of claim 17 is a brain cell.

### Claim 19 (depends on 1)

The cell of claim 1, wherein the mammalian protein complex is a signaling molecule.

### Claim 20 (depends on 19)

The cell of claim 19, wherein the signaling molecule contains a disulfide bond.

### Claim 21 (depends on 20)

The cell of claim 20 is a brain cell.

### Claim 22 (depends on 20)

The cell of claim 20, wherein the fish is a yellow grouper.

### Claim 23 (depends on 22)

The cell of claim 22 is a brain cell.

### Claim 24

A method of expressing in a fish a mammalian protein complex having two peptide species, the method comprising introducing into cells of the fish an eukaryotic expression cassette that contains two sequences encoding a first peptide species and a second peptide species, respectively, of the mammalian protein complex, each sequence being operably linked to a transcription promoter, whereby a multiplicity of peptides of the first peptide species and the second peptide species are expressed and at least one peptide of the first peptide species and at least one peptide of the second peptide species assembly to form the complex.

### Claim 25 (depends on 24)

The method of claim 24, wherein the mammalian protein complex is an antibody.

### Claim 26 (depends on 25)

The method of claim 25, wherein the antibody contains a disulfide bond.

### Claim 27 (depends on 25)

The method of claim 25, wherein the fish is a yellow grouper.

### Claim 28 (depends on 27)

The method of claim 27, wherein the cell is a brain cell.

### Claim 29 (depends on 25)

The method of claim 25, wherein the cell is a brain cell.

### Claim 30 (depends on 25)

The method of claim 25, wherein the antibody binds to a microorganism.

### Claim 31 (depends on 30)

The method of claim 30, wherein the fish is a yellow grouper.

### Claim 32 (depends on 30)

The method of claim 30, wherein the microorganism is a virus.

### Claim 33 (depends on 32)

The method of claim 32, wherein the fish is a yellow grouper.

### Claim 34 (depends on 32)

The method of claim 32, wherein the virus is a nervous necrosis virus.

### Claim 35 (depends on 34)

The method of claim 34, wherein the fish is a yellow grouper.

### Claim 36 (depends on 35)

The method of claim 35, wherein the antibody binds to a coat protein of the nervous necrosis virus.

### Claim 37 (depends on 36)

The method of claim 36, wherein the cell is a brain cell.

### Claim 38 (depends on 24)

The method of claim 24, wherein the mammalian protein complex contains a disulfide bond.

### Claim 39 (depends on 24)

The method of claim 24, wherein the mammalian protein complex is a signaling molecule.

### Claim 40

A cultured fish cell infected with a virus, or a progeny of the cell, wherein the cell produces the virus.

### Claim 41 (depends on 40)

The cell of claim 40, wherein the virus is a yellow grouper nervous necrosis virus.

### Claim 42 (depends on 40)

The cell of claim 40, wherein the cell is a brain cell.

### Claim 43 (depends on 40)

The cell of claim 40, wherein the fish cell is a yellow grouper cell.