Methods and culture media for inducing single cell suspension in insect cell lines
US5728580A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Feb 20, 1996 |
| Grant date | Mar 17, 1998 |
| Priority date | — |
| Expiry date | Feb 20, 2016 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2501/90
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Non-carboxylated sulfated polyanions have been successfully used to rapidly obtain and maintain stable single-cell suspension of BTI-TN5B1-4 cells, a cell line which has a high intrinsic capacity for the expression of recombinant protein, but which clumps severely in suspension reducing its effectiveness as a host for foreign protein production with the baculovirus expression vector system. The three most effective polyanions for inducing a single-cell suspension were dextran sulfate, polyvinyl sulfate, and pentosan sulfate. The cost of dextran sulfate treatment is low compared to heparin treatment, which required a 20-fold higher lever to induce single-cell suspension. More importantly, dextran sulfate does not block vital infection at MOI.gtoreq.1 whereas heparin is known to seriously inhibit infection. To overcome this effect, the cells can be subcultured to a fresh culture medium without the sulfated polyanion, or the sulfated polyanion is neutralized with a polycation, then the cells are inoculated with the baculovirus. Once the cells are infected with the baculovirus, the sulfate polyanion is added back to the fresh culture medium. Inducing single-cell suspension with dextran…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.