Recombinant primate granulocyte macrophage-colony stimulating factor
US5942221A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jun 6, 1995 |
| Grant date | Aug 24, 1999 |
| Priority date | — |
| Expiry date | Jun 6, 2015 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2840/105
- WIPO fieldPharmaceuticals
- WIPO sectorChemistry
Abstract
A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises: preparing RNA from a cell that produces CSF; preparing polyadenylated messenger RNA from said RNA; preparing single stranded cDNA from said messenger RNA; converting the single stranded cDNA to double stranded cDNA; inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies; picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool; transfecting the plasmid DNA into suitable host cells for expressing CSF protein; culturing the transfected cells and assaying the supernatant for CSF activity; and selecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.