# US 6,294,356 B1 · Methods and materials for making and using laminin-5

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- Number: US 6,294,356 B1
- Document ID: US6294356B1
- Status: Granted
- Kind: B1
- Application number: 09/232,394
- Filed: 1999-01-15
- Granted: 2001-09-25
- Earliest claimed: 1998-01-16
- Assignees: Northwestern University
- Inventors: Jonathan C. R. Jones; M. Sharon Stack; Lawrence E. Goldfinger
- Web page: https://patentgrep.com/patent/US6294356B1

## CPC

- C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
- 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 provides two forms of laminin-5. One form comprising unprocessed α3 subunits promotes the migration of epithelial cells. The other form comprising plasmin-cleaved α3 subunits promotes the formation of hemidesmosomes. The invention further provides methods of making and using the two forms of laminin-5 and products comprising the two forms of laminin-5.

## Claims

### Claim 1

A method of generating hemidesmosome-promoting laminin-5 comprising:
culturing epithelial cells that do not produce hemidesmosome-promoting laminin-5 under conditions effective so that they produce extracellular matrix protein comprising laminin-5 containing unprocessed α3 subunits; and
contacting the laminin-5 with plasmin under conditions effective so that the plasmin cleaves the α3 subunits.

### Claim 2 (depends on 1)

The method of claim 1 wherein the plasmin is generated by contacting plasminogen with a plasminogen activator.

### Claim 3 (depends on 2)

The method of claim 2 wherein the plasminogen activator is a tissue-type plasminogen activator.

### Claim 4 (depends on 2)

The method of claim 2 wherein the extracellular matrix protein is contacted with the plasminogen and then with the plasminogen activator.

### Claim 5 (depends on 1)

The method of claim 1 wherein the laminin-5 is human laminin-5.

### Claim 6 (depends on 1)

The method of claim 1 wherein the laminin-5 is isolated from the extracellular matrix protein prior to being contacted with the plasmin.

### Claim 7

A method of generating hemidesmosome-promoting laminin-5 comprising:
producing heterotrimeric laminin-5 containing unprocessed α3 subunits by expressing DNA coding for α3, γ2 and β3 subunits in host cells transformed with the DNA; and
contacting the laminin-5 with plasmin under conditions effective so that the plasmin cleaves the α3 subunits.

### Claim 8 (depends on 7)

The method of claim 7 wherein the α3, γ2 and β3 subunits are produced in the same host cell.

### Claim 9 (depends on 7)

The method of claim 7 wherein the α3, γ2 and β3 subunits are produced in different host cells.

### Claim 10 (depends on 7)

The method of claim 7 wherein the laminin-5 is isolated prior to being contacted with the plasmin.

### Claim 11 (depends on 7)

The method of claim 7 wherein the laminin-5 is human laminin-5.

### Claim 12

A method of generating hemidesmosome-promoting laminin-5 comprising:
contacting a material containing unprocessed α3 subunits of laminin-5, but not γ2 subunits or β3 subunits of laminin-5, with plasmin under conditions effective so that the plasmin cleaves the α3 subunits; and
contacting the cleaved α3 subunits with γ2 and β3 subunits so that the three subunits combine to form the hemidesmosome-promoting laminin-5.

### Claim 13 (depends on 12)

The method of claim 12 wherein the laminin-5 is human laminin-5.

### Claim 14 (depends on 1, 7, 12)

The method of claim 1, 7 or 12 further comprising applying the hemidesmosome-promoting laminin-5 to a shaped article.