# US 5,702,937 A · Method for potentiating tissue plasminogen activator with .beta.-lactoglobulin

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- Number: US 5,702,937 A
- Document ID: US5702937A
- Status: Granted
- Kind: A
- Application number: 08/448,011
- Filed: 1995-05-23
- Granted: 1997-12-30
- Earliest claimed: 1995-05-23
- Assignees: Eli Lilly and Company
- Inventors: Craig M. Zwickl
- Web page: https://patentgrep.com/patent/US5702937A

## CPC

- C12N 9/6459: {t-plasminogen activator (3.4.21.68), i.e. tPA}
- C07K 16/40: against enzymes
- C12Q 1/56: involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
- C12Y 304/21069: Protein C activated (3.4.21.69)
- G01N 33/86: involving blood coagulating time {or factors, or their receptors}
- G01N 2333/9726: Tissue plasminogen activator
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- 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
- C12Y: ENZYMES
- 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)

## Abstract

A method for potentiating the activity of tissue plasminogen activator proteins by adding certain amounts of beta-lactoglobulin A is claimed. In particular, the invention includes an enhanced solid-phase immunoassay for measuring tissue plasminogen activator (nt-PA) activity in a mammalian serum sample with sufficient sensitivity to detect neutralizing activity in samples containing .mu.g/ml levels of total nt-PA antibody. Neutralizing activity can be easily detected in serum dilutions containing as little as 0.038 to 0.48 .mu.g of total nt-PA antibody in a 20 .mu.l sample. The assay has a sensitivity of 1 pM of nt-PA (67 pg/ml) in 5 hr.

## Claims

### Claim 1

A method for increasing the enzymatic activity of a mammalian tissue plasminogen activator protein which comprises adding from 5 to 50 .mu.g/mL of beta lactoglobulin A to a composition containing the mammalian tissue plasminogen activator protein.

### Claim 2 (depends on 1)

The method of claim 1 wherein the tissue plasminogen activator protein is selected from the group consisting of native tissue plasminogen activator and modified tissue plasminogen activator 6 (mt-PA 6).

### Claim 3 (depends on 1)

The method of claim 1 wherein said tissue plasminogen activator protein is native tissue plasminogen activator.

### Claim 4 (depends on 1)

The method of claim 1 wherein said tissue plasminogen activator protein is mt-PA 6.