Mature apoai protein production under serum free culturing conditions
Abstract
Abstract A method of producing a purified lipid-binding peptide which can bind to phospholipids at one or more amphipatic alpha-helical peptide regions. The method includes providing a gene coding for the peptide, and introducing the gene in expressible, heterologous form in a suitable expression system capable of synthesizing a mixture of peptides which includes the lipid-binding peptide. Addition of either endogenous or exogenous lipids to the peptide mixture forms a low-density lipopeptide complex composed of lipid and the lipid-binding peptide, and this complex can be separated easily from nonlipid-binding peptides in the peptide on the basis of its size and/or density. The method is intended particularly for scaled-up production of purified human apolipoproteins and their alpha-helical lipid-binding regions. Also disclosed are related methods for producing recombinant apolipoproteins, therapeutic lipopeptide compositions, and a stabilized lipid emulsion for nutritional therapy. Further disclosed are methods for expressing apolipoproteins or lipid-binding segments thereof in bacterial, yeast and mammalian cell expression systems, and methods for purifying lipid binding proteins, including fused recombinant proteins.
Inventors
- 👤 Andrew A. Protter
- 👤 Jean-Louis Vigne
- 👤 Joanne B. Mallory
- 👤 Karen D. Talmadge
- 👤 John P. Kane
Bibliographic Details
Claims Analysis
Total Claims: 6References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-07-23 | 1994-01-22 | 1994-07-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-07-23 | 1998-01-22 | 1998-07-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-07-23 | 2002-01-22 | 2002-07-24 | $7700.00 | Window Closed / Lapsed |