PURIFICATION AND CHARACTERIZATION OF RECOMBINANT HUMAN MACROPHAGE-COLONY-STIMULATING FACTOR EXPRESSED IN CHINESE-HAMSTER OVARY CELLS
Takahashi, M; Nishida, T; Takano, M; Yamanishi, K; Shimokura, M; Ohmoto, Y; Aihara, K; Ichikawa, H; Nakai, S; Hirai, Y; Adachi, M
| HERO ID | 5410441 |
|---|---|
| In Press | No |
| Year | 1993 |
| Title | PURIFICATION AND CHARACTERIZATION OF RECOMBINANT HUMAN MACROPHAGE-COLONY-STIMULATING FACTOR EXPRESSED IN CHINESE-HAMSTER OVARY CELLS |
| Authors | Takahashi, M; Nishida, T; Takano, M; Yamanishi, K; Shimokura, M; Ohmoto, Y; Aihara, K; Ichikawa, H; Nakai, S; Hirai, Y; Adachi, M |
| Journal | Bioscience, Biotechnology, and Biochemistry |
| Volume | 57 |
| Issue | 6 |
| Page Numbers | 915-921 |
| Abstract | We expressed human macrophage colony-stimulating factor (M-CSF) in Chinese hamster ovary (CHO) cells by introducing an expression plasmid coding for a 554-amino-acid M-CSF precursor and dihydrofolate reductase (DHFR) gene, and by amplifying the sequence. A cell line was obtained that secreted approximately 200,000 units/ml after 6 days in culture. The expressed recombinant human M-CSF (rhM-CSF) primarily consisted of two molecular species, a main 80-90 kD M-CSF as a homodimer and a molecular form higher than 150 kD. Purification of a main rhM-CSF gave an apparently homogeneous protein disulfide-bonded from 42-kD subunits, but one of the purified rhM-CSFs was composed of two subunit species with molecular masses of 44 and 42 kD. These purified rhM-CSFs had substantially the same specific activity (1 to 4 x 10(7) units/mg protein). Deglycosylation experiments with the latter rhM-CSF using chemical (trifluoromethanesulfonic acid) and enzymatic methods found a terminal neuraminic acid in addition to N- and O-glycosylation, but the two subunit species did not coalesce into a single molecule. |
| Doi | 10.1271/bbb.57.915 |
| Pmid | 7763877 |
| Wosid | WOS:A1993LK59400010 |
| Is Certified Translation | No |
| Dupe Override | No |
| Is Public | Yes |
| Language Text | English |