# US 5,707,853 A · Nucleic acid encoding calf intestinal alkaline phosphatase

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- Number: US 5,707,853 A
- Document ID: US5707853A
- Status: Granted
- Kind: A
- Application number: 08/368,071
- Filed: 1995-01-03
- Granted: 1998-01-13
- Earliest claimed: 1992-03-10
- Assignees: La Jolla Cancer Research Foundation
- Inventors: Jose L. Millan
- Web page: https://patentgrep.com/patent/US5707853A

## CPC

- C12Q 1/42: involving phosphatase
- C12N 9/16: acting on ester bonds (3.1)
- G01N 33/535: with enzyme label {or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates}
- C07K 2319/00: Fusion polypeptide
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- 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)

## Abstract

The invention relates to isolated nucleic acids encoding recombinant calf intestinal alkaline phosphatase. Expression vectors and host cells transformed or transfected with such vectors are also provided. The invention further provides multifunctional polypeptides containing amino acid sequences encoding for calf intestinal alkaline phosphatase and a second amino acid sequence encoding a reagent having specific reactivity with a ligand. The recombinant calf intestinal alkaline phosphatase or its active fragments and the multifunctional polypeptides can be used in the methods for determining the presence or concentration of a ligand.

## Claims

### Claim 1

An isolated nucleic acid having a nucleotide sequence as shown in FIG. 1 (SEQ ID NO: 9).

### Claim 2 (depends on 1)

A cDNA encoded by the nucleic acid molecule of claim 1.

### Claim 3 (depends on 1)

An RNA encoded by the nucleic acid molecule of claim 1.

### Claim 4 (depends on 1)

The isolated nucleic acid of claim 1, further comprising a second nucleotide sequence encoding a polypeptide having specific reactivity with a ligand.

### Claim 5 (depends on 1)

A vector comprising the nucleic acid of claim 1.

### Claim 6 (depends on 5)

The vector of claim 5, wherein said vector is a plasmid.

### Claim 7 (depends on 5)

A recombinant host cell comprising the vector of claim 5.

### Claim 8 (depends on 7)

A method of obtaining recombinant calf intestinal alkaline phosphatase comprising culturing said recombinant host cell of claim 7 and isolating said calf intestinal alkaline phosphatase from said culture.

### Claim 9 (depends on 7)

A cell culture comprising the recombinant host cell of claim 7 cultured in a suitable medium.