# US 2013/0157293 A1 · Pharmaceutical manufacturing methods

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- Number: US 2013/0157293 A1
- Document ID: US20130157293A1
- Status: Published application
- Kind: A1
- Application number: 13/605,858
- Filed: 2012-09-06
- Published: 2013-06-20
- Earliest claimed: 2006-08-01
- Assignees: Kineta Two, LLC
- Inventors: MARK E BRANUM; Maralee McVean; Just Justesen; Eric J. Tarcha
- Web page: https://patentgrep.com/patent/US20130157293A1

## CPC

- C12N 9/93: {Ligases (6)}
- C12N 9/1241: {Nucleotidyltransferases (2.7.7)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention describes methods for manufacturing oligoadenylate synthetase (OAS) proteins for use as active pharmaceutical ingredients in pharmaceutical compositions. A manufacturing method is described that produces large quantities of concentrated, highly active OAS protein for use in pharmaceutical compositions for the treatment of a variety of diseases including viral infection. Methods for monitoring and validating the manufacturing process are also described.

## Claims

### Claim 1

A method for producing oligoadenylate synthetase (OAS) proteins comprising:
a) culturing a host cell containing an expression vector in growth medium under conditions wherein the OAS protein is expressed;
b) recovering the host cell from the growth medium; and
c) isolating the OAS protein from the host cell.

### Claim 2 (depends on 1)

The method of claim 1 wherein said OAS protein is selected from the group consisting of SEQ ID NO:9-16.

### Claim 3 (depends on 1)

The method of claim 1 wherein said expression vector comprises a polypeptide selected from the group consisting of SEQ ID NO:1-8.

### Claim 4 (depends on 1)

The method of claim 1 wherein said host cell is E. coli.

### Claim 5 (depends on 1)

The method of claim 1 further comprising a step of assessing in-process protein purity.

### Claim 6 (depends on 5)

The method of claim 5 wherein said step comprises an enzyme-linked immunosorbent assay.

### Claim 7 (depends on 5)

The method of claim 5 wherein said step comprises Limulus Amoebocyte Lysate endotoxin assay.

### Claim 8 (depends on 1)

The method of claim 1 further comprising a step of assessing final purified protein purity.

### Claim 9

A method of purifying an OAS protein comprising:
a) culturing a host cell containing an expression vector in growth medium under conditions wherein the OAS protein is expressed;
b) recovering the host cell from the medium;
c) isolating the OAS protein from the host cell; and
d) purifying the OAS protein using affinity chromatography.

### Claim 10 (depends on 9)

The method of claim 9 wherein said OAS protein comprises an affinity tag.

### Claim 11 (depends on 9)

The method of claim 9 wherein said OAS protein is selected from the group consisting of SEQ ID NO:9-16.

### Claim 12 (depends on 9)

The method of claim 9 wherein said expression vector comprises a polypeptide selected from the group consisting of SEQ ID NO:1-8.

### Claim 13 (depends on 9)

The method of claim 9 wherein said host cell is E. coli.

### Claim 14

A method of purifying an OAS protein comprising:
a) culturing a host cell containing an expression vector in growth medium under conditions wherein the OAS protein is expressed;
b) recovering the host cell from the medium;
c) isolating the OAS protein from the host cell; and
d) purifying the OAS protein using anion exchange chromatography.

### Claim 15 (depends on 14)

The method of claim 14 wherein said OAS protein comprises an affinity tag.

### Claim 16 (depends on 14)

The method of claim 14 wherein said OAS protein is selected from the group consisting of SEQ ID NO:9-16.

### Claim 17 (depends on 14)

The method of claim 14 wherein said expression vector comprises a polypeptide selected from the group consisting of SEQ ID NO:1-8.

### Claim 18 (depends on 14)

The method of claim 14 wherein said host cell is E. coli.