Enhanced production of recombinant proteins by transient transfection of suspension-growing mammalian cells
US10421950B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | May 7, 2002 |
| Grant date | Sep 24, 2019 |
| Priority date | — |
| Expiry date | Jan 19, 2029 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2840/44
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Disclosed is a new process for the production of recombinant proteins, by transient transfection of suspension-grown human embryonic kidney cells (293 cell line and its genetic variants) with an expression vector, using polyethylenimine (PEI) as a transfection reagent. In a preferred embodiment, the process uses 293E cells expressing the Epstein-Barr virus (EBV) EBNA 1 protein, in combination with an oriP-based episomal expression vector having an improved cytomegalovirus expression cassette comprising the CMV5 promoter. The process combines in a single step the cell growth, transfection and protein expression, is carried out without changing the culture medium, and allows to achieve high expression levels in a short period of time. The process may be carried out in a serum-free, low-protein culture medium, is easily scalable, compatible with continuous production processes, and fully adapted to high-throughput production of milligram quantities of recombinant proteins.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.