# US 7,408,053 B1 · Recombinant DNA constructs for GAG polypeptides and polypeptides produced thereby

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- Number: US 7,408,053 B1
- Document ID: US7408053B1
- Status: Granted
- Kind: B1
- Application number: 08/442,750
- Filed: 1995-05-17
- Granted: 2008-08-05
- Earliest claimed: 1984-10-31
- Assignees: Novartis Vaccines and Diagnostics, Inc.
- Inventors: Paul A. Luciw; Dino Dina
- Web page: https://patentgrep.com/patent/US7408053B1

## CPC

- C07K 14/005: from viruses
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/12: Antivirals
- A61P 31/14: for RNA viruses
- A61P 31/18: for HIV
- A61P 37/00: Drugs for immunological or allergic disorders
- A61P 37/02: Immunomodulators
- C12N 9/0089: {acting on superoxide as acceptor (1.15)}
- C12N 15/81: for yeasts
- C12N 15/85: for animal cells
- C12Q 1/703: {Viruses associated with AIDS}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 2319/00: Fusion polypeptide
- C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation
- C07K 2319/735: containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
- C07K 2319/75: containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
- C12N 2740/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2740/16222: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2800/108: episomal vectors
- C12N 2830/00: Vector systems having a special element relevant for transcription
- C12N 2830/002: inducible enhancer/promoter combination, e.g. hypoxia, iron, transcription factor
- C12N 2830/005: repressible enhancer/promoter combination, e.g. KRAB
- C12N 2830/15: chimeric enhancer/promoter combination
- C12N 2830/36: being a transcription termination element
- C12N 2830/55: from bacteria
- C12N 2830/702: yeast
- C12N 2830/704: S. cerevisiae
- Y10S 435/81: Packaged device or kit
- Y10S 435/82: Subcellular parts of microorganisms
- Y10S 435/948: using viruses or cell lines
- Y10S 435/974: Aids related test
- Y10S 435/975: Kit
- Y10S 530/826: Viruses
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Polynucleotide sequences are provided for the diagnosis of the presence of retroviral infection in a human host associated with lymphadenopathy syndrome and/or acquired immune deficiency syndrome, for expression of polypeptides and use of the polypeptides to prepare antibodies, where both the polypeptides and antibodies may be employed as diagnostic reagents or in therapy, e.g., vaccines and passive immunization. The sequences provide detection of the viral infectious agents associated with the indicated syndromes and can be used for expression of antigenic polypeptides.

## Claims

### Claim 1

A recombinant DNA construct useful for the expression of a recombinant polypeptide in a cell containing the construct, the construct comprising control sequences which regulate transcription and translation of the recombinant polypeptide in the cell and a coding sequence regulated by the control sequences, wherein the coding sequence comprises a DNA sequence of at least about 27 bp of the gag open reading frame shown in FIG. 4, characterized in that the DNA sequence encodes an antigenic gag HIV amino acid sequence which amino acid sequence is immunologically non-cross-reactive with HTLV-I and HTLV-II.

### Claim 2 (depends on 1)

A recombinant DNA construct according to claim 1 which comprises a replication system functional in a eukaryotic cell.

### Claim 3 (depends on 1)

A recombinant DNA construct according to claim 1 which comprises a replication system functional in a yeast cell.

### Claim 4 (depends on 1)

A recombinant DNA construct according to claim 1 which comprises a replication system functional in a bacterial cell.

### Claim 5 (depends on 1)

A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises p18 gag.

### Claim 6 (depends on 1)

A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises p25 gag.

### Claim 7 (depends on 1)

A recombinant DNA construct according to claim 1 wherein the antigenic amino acid sequence comprises a fusion protein of p18 gag and p25 gag.

### Claim 8 (depends on 1, 2, 3, 4)

A recombinant DNA construct according to any one of claims 1-4 wherein the antigenic amino acid sequence comprises a complete gag sequence.

### Claim 9 (depends on 1, 2, 3, 4, 5, 6)

A cell comprising a recombinant DNA construct according to any one of claims 1 to 4, 5, 6, or 7, wherein the cell expresses the antigenic HIV amino acid sequence and is free from other cells which do not express the antigenic HIV amino acid sequence.

### Claim 10 (depends on 2)

A recombinant antigenic gag HIV polypeptide produced by the construct of claim 2 expressed in an eukaryotic cell.

### Claim 11 (depends on 3)

A recombinant antigenic gag HIV polypeptide produced by the construct of claim 3 expressed in a yeast cell.

### Claim 12 (depends on 4)

A recombinant antigenic gag HIV polypeptide produced by the construct of claim 4 expressed in a bacterial cell.

### Claim 13 (depends on 5)

A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 5.

### Claim 14 (depends on 6)

A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 6.

### Claim 15 (depends on 7)

A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 7.

### Claim 16 (depends on 8)

A recombinant antigenic gag HIV polypeptide produced by the DNA construct of claim 8.

### Claim 17

A purified polynucleotide of at least about 27 bp of the gag open reading frame shown in FIG. 4 which encodes an antigenic gag HIV amino acid sequence which amino acid sequence is immunologically non-cross-reactive with HTLV-1 and HTLV-II.

### Claim 18 (depends on 17)

The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises p18 gag.

### Claim 19 (depends on 17)

The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises p25 gag.

### Claim 20 (depends on 17)

The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises a fusion protein of p18 gag and p25 gag.

### Claim 21 (depends on 17)

The polynucleotide of claim 17 wherein the antigenic amino acid sequence comprises a complete gag sequence.