Inducing single-cell suspension of BTI-TN5B1-4 insect cells: II. The effect of sulfated polyanions on baculovirus infection

✍ Scribed by Kennie U. Dee; H. Alan Wood; Michael L. Shuler

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 437 KB
• Volume: 54
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19970505)54:3<206::aid-bit2>3.0.co;2-j

✦ Synopsis

Sulfated polyanions can be used to rapidly in-hancement of recombinant protein production (Dee, duce and maintain single-cell suspensions of BTIet al. 1997b). The enhancement may be a combined TN5B1-4 insect cells, a cell line which clumps in suspenresult of improved nutrient and oxygen transport, insion. Elimination of cell clumping results in a significant creased accessibility of the cells to the virus, and eliminaincrease in volumetric yield of the baculovirus expression tion of cell-cell contact which is known to severely invector system. Sulfated polyanions, however, inhibited baculovirus infection of BTI-TN5B1-4. Data from binding hibit protein synthesis in attached cultures (Chung et al., studies and fusion assays suggest that the inhibition of 1993; Wickham, 1991).

infection was not due to the observed reduction in viral Sulfated polysaccharides, however, are known to poattachment rate but to inhibition of viral membrane fusion tently inhibit in vitro replication of a large number of in the endosome. enveloped viruses including members of the toga-, The three most effective polyanions for inducing single cells are dextran sulfate, pentosan sulfate, and polyvinyl herpes-, paramyxo-, orthomyxo-, rhabdo-, arena-, and sulfate. At concentrations required for single-cell formaretrovirus family (Krumbiegel et al., 1992; Luschertion, dextran sulfate and pentosan sulfate did not affect