Production of recombinant proteins utilizing non-antibiotic selection methods and the incorporation of non-natural amino acids therein
US9243253B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 10, 2010 |
| Grant date | Jan 26, 2016 |
| Priority date | — |
| Expiry date | Feb 12, 2033 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/78
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.