Methods for expression and purification of recombinant human growth hormone

• US 8,178,108 B2
• Filed: 10/17/2007
• Issued: 05/15/2012
• Est. Priority Date: 12/22/2004
• Status: Expired due to Fees

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: Buechler, Ying, Lieu, Ricky, Ong, Michael, Bussell, Stuart, Knudsen, Nick, Cho, Ho Sung
• Primary Examiner(s): Stoica, Elly-Gerald
• Application Number:
  US11/874,086
• Publication Number:
  US 20080050777A1
• Time in Patent Office: 1,672 Days
• Field of Search: None
• US Class Current: 424/198.1
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin

Abstract

The present invention relates generally to the production, purification, and isolation of human growth hormone (hGH). More particularly, the invention relates to the production, purification, and isolation of substantially purified hGH comprising non-naturally encoded amino acids and hGH from recombinant host cells or culture medium including, for example, yeast, insect, mammalian and bacterial host cells. The process of the present invention is also useful for purification of hGH linked to polymers or other molecules.

Claims

1. A process of producing water soluble polymer-human growth hormone (hGH) polypeptide conjugate with a serum half-life at least two-fold greater than the serum half-life of a naturally occurring, unconjugated version of an hGH protein, comprising:

a) culturing recombinant host cells capable of expressing an hGH polypeptide comprising a non-naturally encoded amino acid having the structure;

2. The process of claim 1, wherein the water soluble polymer is polyethylene glycol.

3. The process of claim 1 further comprising h) contacting the hGH conjugate from step g) with a hydrophobic interaction chromatography (HIC) matrix under conditions that allow binding of the hGH to the matrix;

and i) eluting and collecting the hGH conjugate from the HIC matrix to provide substantially pure hGH conjugate comprising a non-naturally encoded amino acid.

4. The process of claim 1, wherein said recombinant host cell is a eukaryotic cell.

5. The process of claim 1, wherein said recombinant host cell is a prokaryotic cell.