Establishment of a large-scale purification procedure for purified recombinant bovine interferon-tau produced by a silkworm-baculovirus gene expression system
| Autor: | Yuichi Yokomizo, Hiroki Kaki, Hitomi Takahashi, Masashi Takahashi, Hidekazu Nagaya, Shigeki Inumaru, Toshimichi Kanaya, Yoneko Tobita |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Genetic Vectors
Gene Expression Biology Pregnancy Proteins Recombinant virus Neutralization law.invention Sepharose chemistry.chemical_compound Column chromatography law Complementary DNA Gene expression Animals Chromatography General Veterinary Silica gel fungi Hydrogen-Ion Concentration Bombyx Molecular biology Recombinant Proteins chemistry Larva Interferon Type I Recombinant DNA Cattle Baculoviridae Biotechnology |
| Zdroj: | The Journal of veterinary medical science. 66(11) |
| ISSN: | 0916-7250 |
| Popis: | We developed a procedure for the large-scale purification of bovine interferon-tau (boIFN-tau) by means of a silkworm-baculovirus gene expression system. Recombinant boIFN-tau (rboIFN-tau) was efficiently produced in the silkworm infected with boIFN-tau cDNA recombinant baculovirus and accumulated in the haemolymph. To establish a purification method suitable for mass production, we tried three crude purification methods, namely, an acidification and neutralization treatment (ANT), silica gel column chromatography (SGCC), and Blue sepharose column chromatography (BSCC) with a combination of Q-sepharose (QSC) and chelating sepharose column chromatographies (CSCC). As a result, the acidification and neutralization treatment was found to be the most efficient and cost effective. With this combination, we obtained 91% pure products. To confirm the applicability of the procedure for mass production, we inoculated 100 silkworms with the recombinant virus, and recovered about 4.55 mg (1.26 x 10(8)U/mg) of 91% pure rboIFN-tau by means of a combination of the ANT, followed by QSC and CSCC. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|