Safety aspects related to recombinant protein expression from Semliki Forest virus vectors |
| |
Authors: | Kenneth Lundstrom Daniel Rotmann Danielle Hermann Ernst-Jürgen Schlaeger |
| |
Institution: | (1) Research Laboratories, F. Hoffmann-La Roche, CH-4070 Basel, Switzerland;(2) Research Laboratories, F. Hoffmann-La Roche, CH-4070 Basel, Switzerland |
| |
Abstract: | Semliki Forest virus vectors (SFV) have been developed for efficient transgene expression to result in high receptor yields(50–200
pmol receptor/mg protein) in a variety of mammalian host cells. Transfer of the SFV technology to mammalian cells growing
in suspension cultures has made it feasible to produce hundreds of milligrams of receptor proteins in a short time. Large-scale
production, however, raises the questions of the safety of handling virally infected cells for down-stream processing. Analysis
of cell culture medium and SFV-infected cells revealed that some infectious particles were still present. Replacement of virus-containing
medium at 2 h post-infection efficiently removed the majority of infectious replication-deficient SFV particles. Washes with
PBS further reduced the number of infectious particles significantly both in the medium and associated with cells to levels
that allowed safe handling of SFV-infected cells outside the cell culture facility for biochemical, pharmacological, or electrophysiological
assays or down-stream processes in connection to receptor purification. Furthermore, engineering of novel temperature-sensitive
mutant SFV vectors resulted in temperature-controlled transgene expression, which completely eliminates the risk of contaminating
laboratory personnel.
This revised version was published online in July 2006 with corrections to the Cover Date. |
| |
Keywords: | mutant vectors recombinant proteins safety aspects Semliki Forest virus |
本文献已被 PubMed SpringerLink 等数据库收录! |
|