# US 5,708,155 A · Enhanced immunogenicity using leukotoxin chimeras

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- Number: US 5,708,155 A
- Document ID: US5708155A
- Status: Granted
- Kind: A
- Application number: 08/455,970
- Filed: 1995-05-31
- Granted: 1998-01-13
- Earliest claimed: 1991-10-16
- Assignees: University of Saskatchewan
- Inventors: Andrew A. Potter; Mark J. Redmond; Huw P. A. Hughes
- Web page: https://patentgrep.com/patent/US5708155A

## CPC

- C07K 14/005: from viruses
- A61K 39/0006: {Contraceptive vaccins; Vaccines against sex hormones}
- A61K 47/62: the modifying agent being a protein, peptide or polyamino acid
- A61K 47/646: {the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines}
- A61K 47/6829: {Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A}
- A61K 47/6833: {Viral toxins}
- C07K 5/1019: {with the first amino acid being basic}
- C07K 5/1021: {with the first amino acid being acidic}
- C07K 5/1024: {with the first amino acid being heterocyclic}
- C07K 7/23: Luteinising hormone-releasing hormone [LHRH]; Related peptides
- C07K 14/285: from Pasteurellaceae (F), e.g. Haemophilus influenza
- C07K 14/655: Somatostatins
- C07K 16/26: against hormones {; against hormone releasing or inhibiting factors}
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- A61K 2039/6037: {Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]}
- C07K 2319/00: Fusion polypeptide
- C12N 2720/12322: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

New immunological carrier systems, DNA encoding the same, and the use of these systems, are disclosed. The carrier systems include chimeric proteins which comprise a leukotoxin polypeptide fused to a selected antigen. The leukotoxin functions to increase the immunogenicity of the antigen fused thereto.

## Claims

### Claim 1

A DNA construct encoding a chimeric protein, said DNA construct comprising a first nucleotide sequence encoding a leukotoxin polypeptide capable of activating helper T-cells directed to a selected antigen, operably linked to a second nucleotide sequence encoding said selected antigen.

### Claim 2 (depends on 1)

The DNA construct of claim 1 wherein said second nucleotide sequence encodes somatostatin (SRIF), or an epitope thereof.

### Claim 3 (depends on 2)

The DNA construct of claim 2 comprising the nucleotide sequence depicted in SEQ ID NO:9.

### Claim 4 (depends on 1)

The DNA construct of claim 1 wherein said second nucleotide sequence encodes gonadotropin releasing hormone (GnRH) comprising the amino acid sequence Gln-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly, or an epitope thereof.

### Claim 5 (depends on 4)

The DNA construct of claim 4 comprising the nucleotide sequence depicted in SEQ ID NO:11.

### Claim 6 (depends on 1)

The DNA construct of claim 1 wherein said second nucleotide sequence encodes bovine rotavirus VP4, or an epitope thereof.

### Claim 7 (depends on 6)

The DNA construct of claim 6 comprising the nucleotide sequence depicted in SEQ ID NO:13.

### Claim 8 (depends on 1)

An expression cassette comprised of:
(a) the DNA construct of claim 1; and
(b) control sequences that direct the transcription of said construct whereby said construct can be transcribed and translated in a host cell.

### Claim 9 (depends on 2)

An expression cassette comprised of:
(a) the DNA construct of claim 2; and
(b) control sequences that direct the transcription said construct whereby said construct can be transcribed and translated in a host cell.

### Claim 10 (depends on 4)

An expression cassette comprised of:
(a) the DNA construct of claim 4; and
(b) control sequences that direct the transcription said construct whereby said construct can be transcribed and translated in a host cell.

### Claim 11 (depends on 6)

An expression cassette comprised of:
(a) the DNA construct of claim 6; and
(b) control sequences that direct the transcription said construct whereby said construct can be transcribed and translated in a host cell.

### Claim 12 (depends on 8)

A host cell stably transformed with the expression cassette of claim 8.

### Claim 13 (depends on 9)

A host cell stably transformed with the expression cassette of claim 9.

### Claim 14 (depends on 10)

A host cell stably transformed with the plasmid of claim 10.

### Claim 15 (depends on 11)

A host cell stably transformed with the plasmid of claim 11.

### Claim 16 (depends on 12)

A method of producing a recombinant polypeptide comprising:
(a) providing a population of host cells according to claim 12; and
(b) growing said population of cells under conditions whereby the polypeptide encoded by said expression cassette is expressed.

### Claim 17 (depends on 13)

A method of producing a recombinant polypeptide comprising:
(a) providing a population of host cells according to claim 13; and
(b) growing said population of cells under conditions whereby the polypeptide encoded by said expression cassette is expressed.

### Claim 18 (depends on 14)

A method of producing a recombinant polypeptide comprising:
(a) providing a population of host cells according to claim 14; and
(b) growing said population of cells under conditions whereby the polypeptide encoded by said expression cassette is expressed.

### Claim 19 (depends on 15)

A method of producing a recombinant polypeptide comprising:
(a) providing a population of host cells according to claim 15; and
(b) growing said population of cells under conditions whereby the polypeptide encoded by said expression cassette is expressed.